某皂苷类样品

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍bioGenousTMMouse Intestinal Organoid Kit（小鼠小肠类器官试剂盒）是一款用于建立和维持小鼠肠道成体干细胞来源小肠类器官的完全培养基。生长在该完全培养基中的小鼠小肠类器官主要由干细胞、肠祖细胞和肠绒毛细胞、潘氏细胞、杯状细胞和少量肠内分泌细胞组成，在自我更新能力、组织结构、细胞类型和功能方面，小鼠小肠类器官重现了体内小肠上皮的特征，因此是肠道稳态和疾病机制研究的理想体外模型。技术参数产品信息:产品组成货号体积储存条件及周期bioGenousTMMouse Intestinal Organoid Basal MediumK2001-MI-A100/A500100mL/500mL4℃，12个月bioGenousTMMouse Intestinal Organoid Supplement B(50x)K2001-MI-B100/B5002mL/10mL-20℃,避免反复冻融，12个月bioGenousTMMouse IntestinalOrganoid Supplement C(250x)K2001-MI-C100/C5000.4mL/2mL-20℃,避免反复冻融，12个月EDTA (0.5M, pH 8.0)E2191210.2mL/1mL15-30℃，5年小鼠小肠类器官完全培养基的制备：使用无菌操作技术配制小鼠小肠类器官完全培养基。以下是准备10mL完全培养基的示例，如所需量不同，可相应调整用量。1.冰上解冻Mouse Intestinal Organoid Supplement B (50x)和Mouse Intestinal Organoid Supplement C (250x)。注意：解冻后，建议将Mouse Intestinal Organoid Supplement B (50x)和Mouse Intestinal Organoid Supplement C (250x)分别分装后保存取用，避免反复冻融。2.将200uLMouse Intestinal Organoid Supplement B (50x)，40uLMouse Intestinal Organoid Supplement C (250x)加至9.76mLMouse Intestinal Organoid Basal Medium中，充分混合，配制成10mL小鼠小肠类器官完全培养基。注意：配制后的小鼠小肠类器官完全培养基可在2-8°C储存，建议在两周内使用。Mouse Intestinal Organoid Supplement B(50x)内含有细菌及真菌抗生素(50x)。小鼠小肠类器官原代培养1.依据所在单位批准的实验动物伦理及操作规范牺牲小鼠。2.准备若干培养皿，加入4℃预冷的DPBS备用。3.标准手术操作取小鼠小肠，根据实验需求取总长度约3厘米至20厘米的小肠段，置于含DPBS的培养皿中。4.使用移液管或者注射器往小肠一端注入DPBS以冲洗肠内容物，冲洗后置于新的含DPBS的培养皿中，重复冲洗数次至内容物完全被冲洗干净，置于新的含DPBS的培养皿中。5.使用手术剪将肠管剪开，肠腔面朝上，一只手使用手术镊夹住肠组织一端，另一只手使用手术刀片轻轻刮去肠腔表面肠绒毛，待肠绒毛被刮净后，将肠组织置于新的含DPBS的培养皿中清洗，重复清洗一次。6.将清洗后的小肠组织剪碎至2mm宽，并转移至含有5mmol/L EDTA的预冷DPBS中消化，置于4℃孵育30min。7.消化完成后，将组织碎片转移到新的含DPBS的培养皿中清洗，重复一次以去除EDTA。8.用5mL移液管在含冷的DPBS的培养皿或50mL离心管中吹打、重悬组织碎片，使组织反复穿过移液管尖以产生机械剪切力从而使隐窝与基底层分离，取一部分悬液镜检，当可以看到大量的隐窝样结构后，停止吹打，并对吹打后的组织悬液进行70μm滤网过滤。9.收集穿过滤网的组织悬液，150g离心力4℃离心3min。10.弃上清，使用1mLDPBS重悬组织沉淀，取20uL悬液进行镜检和隐窝计数，计数完成后吸取包含所需隐窝量的悬液，150g离心力4℃离心3min，弃上清后置于冰上。11.用适量的Matrigel重悬组织沉淀，推荐重悬密度为每10uLMatrigel悬液包含200至600个隐窝，重悬后置于冰上，重悬时间不超过30s以避免Matrigel过早凝固。注意：Matrigel稀释比例应在70%以上以保证培养过程中Matrigel的结构稳定性。12.将Matrigel和组织细胞的混合悬液点入24孔板底部正中央，每孔30uL左右，避免悬液接触孔板侧壁。注意：为防止Matrigel室温凝固，此步骤应尽快完成。13.将接种完成后的培养板至于37℃二氧化碳恒温培养箱中，孵育15min左右待Matrigel凝固。14.配制小鼠小肠类器官完全培养基。15.待Matrigel完全凝固后，加入已配制好的小鼠小肠类器官完全培养基，24孔板每孔500uL。注意：请沿壁缓慢加入，避免破坏已凝固结构。16.将24孔板置于37℃二氧化碳培养箱中培养。17.每3天更换一次培养基，更换培养基时应避免破坏Matrigel。18.密切监测类器官生长状态，理想情况下，小鼠小肠类器官应在5至7天内建成。小鼠小肠类器官传代培养19.用经过Anti-Adherence Rinsing Kit(E238002)润洗的枪头吹打刮取类器官，并将类器官和培养基的悬液转移至经过Anti-Adherence Rinsing Kit润洗的1.5mL EP管中。20.用经过Anti-Adherence Rinsing Kit润洗的枪头用力重悬类器官悬浮液，使得类器官与Matrigel分离。21.150g离心力4℃离心3min，弃上清，使用DPBS重悬底部类器官沉淀，150g离心力4℃再次离心3min，弃上清后置于冰上。22.用适量的Matrigel重悬类器官沉淀，重悬后置于冰上，重悬时间不超过30s以避免Matrigel过早凝固。注意：Matrigel稀释比例应在70%以上以保证培养过程中Matrigel的结构稳定性。23.将Matrigel和类器官的混合悬液点入24孔板底部正中央，避免悬液接触孔板侧壁，每孔30uL左右。注意：为防止Matrigel室温凝固，此步骤应尽快完成。24.将接种完成后的培养板至于37℃二氧化碳恒温培养箱中，孵育15min左右待Matrigel凝固。25.配制小鼠小肠类器官完全培养基。26.待Matrigel完全凝固后，加入已配制好的小鼠小肠类器官完全培养基，24孔板每孔500uL。27.将24孔板置于37℃二氧化碳培养箱中培养。

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMMouse Liver Ductal Organoid Kit is a chemically defined cell culture medium for establishment and maintenance of Mouse ductal organoids(mLDs) derived from adult stem cells. Self-renewal of the ductal epithelium is driven by the proliferation of stem cells and their progenitors located in liver. mLDs display all hallmarks of the ductal epithelium in terms of architecture, cell type composition, and self-renewal dynamics, therefore hold great promise for unprecedented studies of Mouse liver development and disease, mLDs may also have applications in regenerative biology through ex vivo expansion of the ductal epithelia.技术参数Product Information:ComponentComponent Cat#VolumeStorage& StabilitybioGenousTMMouse Liver Ductal Organoid Basal Medium（Expansion）K2006-MLD -A100/A500100mL/500mL4℃，12 monthsbioGenousTMMouse Liver Ductal Organoid Supplement B(50x)（Expansion）K2006-MLD –B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMMouse Liver Ductal Organoid Supplement C(250x)（Expansion）K2006-MLD –C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Mouse Liver Ductal Organoid Expansion Medium and Maintenance MediumUse sterile technique to prepare the mouse liver ductal organoid expansion medium and maintenance medium. mLDs grown in Mouse Liver Ductal Organoid Expansion Medium overwhelmingly consisted of cholangiocytes. The following examples are for preparing 10 mL of Expansion Medium and Maintenance Medium. If preparing other volumes, adjust accordingly.1.Thaw Mouse Liver Ductal Organoid Supplement B(50x) (Expansion), Mouse Liver Ductal Organoid Supplement C(250x) (Expansion) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.For Mouse Liver Ductal Organoid Expansion Medium. Add 200 μL Mouse Liver Ductal Organoid Supplement B(50x)(Expansion), 40 μL Mouse Liver Ductal Organoid Supplement C(250x) (Expansion) to 10 mL Mouse Liver Ductal Organoid Basal Medium (Expansion). Mix thoroughly.NOTE:If not use immediately, store complete medium at 2-8°C for not more than 2 weeks. bioGenousTMMouse Liver Ductal Organoid Supplement B (Expansion) contains fungicide and antibiotics(50x).Protocol for Establishment of Mouse Liver Ductal OrganoidsEstablishment of Organoids from Primary Tissue1.Collect primary Mouse liver tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Rinse the liver tissuewith Epithelial Organoid Basal Medium(B213151) or DPBSuntil the supernatant is clear.3.Before ducts isolation, thaw Matrigel on ice and keep it cold. Add 5 mL of FBS to 45 mL of Epithelial Organoid Basal Medium to prepare 10% (vol/vol) FBS medium.4.Mince the tissue into small fragments of 5 mm3in a cell culture dish using surgical scissors or scalpels.CRITICALThe dissected samples must be small enough to pass through the tip of a 10 mL pipette.5.Place the dissected pieces of sample into a 15 mL concial tube containing 10 mL of cold Epithelial Organoid Basal Medium with 1% FBS.6.Wash the samples by pipetting with a 10 mL pipette at least ten times.CRITICALFor the subsequent steps, coat the inner surface of every 10 mL pipette with 10% (vol/vol) FBS medium before use to avoid adherence of the samples on the pipette wall.7.Stand the tube still until the samples settle at the bottom. Aspirate the supernatant with a 10 mL pipette and add 10 mL of pre-warmed Tissue Digestion Solution (K601008).8.Digest the tissue fractions in 37℃with rotation at the speed of 200 rpm. The digestion time should not exceed 30 mins.CRITICALTo prevent over-digestion, one should examine under the microscope if the duct structure appear during digestion.9.Once the duct structure appears, stop digestion by transfer the digestion solution into 50mL tube and add 40 ml Epithelial Organoid Basal Medium with 1% FBS.10.Stand the tube for 2 min. Transfer the supernatant into a new tube.11.Spin the supernatant at 4°C at 300g for 3 min. Aspirate and discard the supernatant.12.Re-suspend the pellet with 10 ml Epithelial Organoid Basal Medium with 1%FBS, and spin at 4°C at 300g for 3 min.13.Repeat last step for 2 times.14.Aspirate the supernatant and resuspend the pellet in Matrigel. Matrigel should be kept on ice to prevent it from solidifying thus, work quickly. The amount of Matrigel depends on the size of the pellet. Approximately 20-100 ducts should be plated in 25 μL of Matrigel.CRITICALDo not dilute Matrigel too much (Matrigel should be 70% (Matrigel vol/Total vol)) to ensure formation of solid droplets.15.Plate the Matrigel containing organoids on the bottom of 24-well cell culture plates in droplets of ~30 μL each around the center of the well.CRITICALOnce the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.16.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 minto let the Matrigel solidify.17.Prepare the required amount of bioGenousTMMouse Liver Ductal Organoid Expansion Medium.18.Once the Matrigel droplets are solidified (15-25 min), open the plate and carefully add 500 μL of Organoid Complete Medium to each well.CRITICALDo not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.19.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.20.Change the medium every 3 days by carefully aspirating medium from the wells and replacing it with fresh, pre-warmed Organoid Complete Medium.21.Closely monitor the organoid formation. Ideally, Mouse Liver Ductal Organoids should be passaged for the first time between 5 and 8 days after initial plating.Splitting and Passaging of Organoids22.Pipette up and down to disrupt the Matrigel, and transfer the organoid suspension into a 1.5 mL conical tube.23.Pipette the organoid suspension up and down to mix thoroughly.Use the bottom of the tube to create pressure, which will aid the removal of Matrigel.24.Centrifuge organoids at 200g for 3 min at room temperature.25.Aspirate the supernatant, and split organoids using either mechanical disruption or Organoid Dissociation Solution (E238001). For mechanical disruption, resuspend the pellet in 1 mL of Organoid Basal Medium. Use a pipette tip to pipette the organoid suspension up and down 30 times. Use the bottom of the tube to create pressure, which will aid organoid disruption. In case of Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥10 times every 1 min to aid in the disruption of the organoids. Monitor digestion closely to keep the incubation time inOrganoid Dissociation Solution to a minimum.CRITICALDo not dissociate in Organoid Dissociation Solution for 3 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.26.After shearing is complete, wash once by topping up with 1 mL ofOrganoid Basal Medium, and centrifuge at 200g for 3 min at room temperature.27.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets on the bottom of a culture plate as describedin Steps 12. After plating is complete, transfer the plate into ahumidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.28.Pre-warm Mouse Liver Ductal Organoid Maintenance Medium at 37 °C.29.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.30.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMMouse Liver Ductal Organoid Kit is a chemically defined cell culture medium for establishment and maintenance of Mouse ductal organoids(mLDs) derived from adult stem cells. Self-renewal of the ductal epithelium is driven by the proliferation of stem cells and their progenitors located in liver. mLDs display all hallmarks of the ductal epithelium in terms of architecture, cell type composition, and self-renewal dynamics, therefore hold great promise for unprecedented studies of Mouse liver development and disease, mLDs could differentiate into hepatocyte like cell under the induction of differentiation medium.技术参数Product Information:ComponentComponent Cat#VolumeStorage& StabilitybioGenousTMMouse Liver Ductal Organoid Basal Medium（Differentiation）K2006-MLH –A100/A500100mL/500mL4℃，12 monthsbioGenousTMMouse Liver Ductal Organoid Supplement B(50x)（Differentiation）K2006-MLH –B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMMouse Liver Ductal Organoid Supplement C(250x)（Differentiation）K2006-MLH –C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMMouse Liver Ductal Organoid Supplement D(250x)（Differentiation）K2006-MLH –D100/D5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Mouse Liver Ductal Organoid Differentiation MediumUse sterile technique to prepare the mouse liver ductal organoid differentiation medium. mLDs grown in Mouse Liver Ductal Organoid Expansion Medium overwhelmingly consisted of cholangiocytes. After changing the Expansion Medium to differentiation medium, the mLDs could differentiate into hepatocyte like cells, which display the markers of hepatocytes, including Albumin, Ttr, Cyp3a11 and Mup20. The following examples are for preparing 10 mL of Differentiation I Medium and Differentiation II Medium. If preparing other volumes, adjust accordingly.1.Thaw Mouse Liver Ductal Organoid Supplement B(50x) (Differentiation), Mouse Liver Ductal Organoid Supplement C(250x) (Differentiation) and Mouse Liver Ductal Organoid Supplement D(250x) (Differentiation) on ice.NOTE:Once thawed, use immediately or aliquot and store at -20°C for no more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.For Mouse Liver Ductal Organoid Differentiation Medium I. Add 200 μL Mouse Liver Ductal Organoid Supplement B(50x)(Differentiation), 40 μL Mouse Liver Ductal Organoid Supplement C(250x)(Differentiation) to 10 mL Mouse Liver Ductal Organoid Basal Medium (Differentiation). Mix thoroughly.3.For Mouse Liver Ductal Organoid Differentiation Medium II. Add 200 μL Mouse Liver Ductal Organoid Supplement B(50x)（Differentiation）and 40 μL Mouse Liver Ductal Organoid Supplement C(250x)(Differentiation) and 10 μL Mouse Liver Ductal Organoid Supplement D (250x)(Differentiation) to 10 mL Mouse Liver Ductal Organoid Basal Medium (Differentiation). Mix thoroughly.NOTE:If not use immediately, store complete medium at 2-8°C for no more than 2 weeks. bioGenousTMMouse Liver Ductal Organoid Supplement B (Differentiation) contains fungicide and antibiotics(50x).Protocol for Mouse Liver Ductal Organoids Differentiation1.Culture the mouse ductal organoids in Mouse Liver Ductal Organoid Medium(Expansion) (K2006-MLD) for at least 3 days.2.Change the medium to Mouse Liver Ductal Organoid Differentiation Medium I, and culture for 9 days. During this period, replace the medium every day.3.Change the medium to Mouse Liver Ductal Organoid Differentiation Medium II, and culture for 3 days. During this period, replace the medium every day.4.At the end of this period, the differentiation process is completed. The ductal organoid would differentiate into hepatocyte-like organoid with the expression of hepatic markers, such as Albumin, Ttr, Cyp3a11 and Mup20.

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMLung Squamous Carcinoma Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human lung squamous carcinoma organoids. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMLung Squamous Carcinoma Organoid Basal MediumK2140-LS-A100/A500100mL/500mL4℃，12 monthsbioGenousTMLung Squamous Carcinoma Organoid Supplement B (50x)K2140-LS-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMLung Squamous Carcinoma Organoid Supplement C (250x)K2140-LS-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Lung Squamous Carcinoma Complete MediumUse a sterile technique to prepare the lung squamous carcinoma organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1.Thaw Lung Squamous Carcinoma Organoid Supplement B(50x) and Lung Squamous Carcinoma Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL Lung Squamous Carcinoma Organoid Supplement B(50x) and 40μL Lung Squamous Carcinoma Organoid Supplement C(250x) to 9.76mL Lung Squamous Carcinoma Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The Lung Squamous Carcinoma Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived Lung Squamous Carcinoma OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human lung squamous carcinoma tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution (K601003) in a 15mL conical tube at 37°C, with variable incubation times ranging from 30 min to 1.5 h. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestionmixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010) to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well..CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 minto let the Matrigel solidify.12.Prepare the required amount of lung squamous carcinoma organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived lung squamous carcinoma organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥8 times every 2 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 7 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm lung squamous carcinoma organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.25.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

HAB现在被视为一种公共威胁，而水资源管理人员需要可靠的工具 来进行监控. YSI的总藻类传感器是最敏感的工具，帮助您监测、缓解和管理有害藻华的影响. 我们的双通道传感器可以帮助您: 尽早发现藻类的爆发，以规划你的响应 为其他实验室分析收集样品的支持的合理方法，节省你的时间和金钱 了解你系统中藻类生长的季节和天气相关模式 了解其他参数如pH值、溶解氧和温度的相关性总藻传感器规格叶绿素(所有传感器)范 围0- 100RFU或0-400µ g/Lchl精 度线性: r2 ≥ 0.999对罗丹明WT全范围分辨率0.01RFU或0.01µ g/L chl藻青蛋白(TAL-PC传感器）范 围0-100RFU或0-100µ g/LPC精 度线性: r2 ≥ 0.999对罗丹明WT全范围分辨率0.01RFU或0.01µ g/LPC藻红蛋白(TAL-PE传感器）范 围0- 100RFU或0-280µ g/LPE精 度线性 1 : r2 ≥ 0.999对罗丹明WT全范围分辨率0.01RFU或 0.01µ g/LPE附加规格使用温度-5-50°C存储温度-20- 80°C深度等级100m(ProDSS),250m(EXO)保修1年兼容ProDSS,EXO订购信息(分开订购)626210总藻-藻青蛋白(TAL-PC)对ProDSS626211总藻-藻红蛋白(TAL-PE)对ProDSS

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMHead and Neck Squamous Cell Carcinoma Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human head and neck squamous cell carcinoma organoids. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMHead and Neck Squamous Cell Carcinoma Organoid Basal MediumK2109-HN-A100/A500100mL/500mL4℃，12 monthsbioGenousTMHead and Neck Squamous Cell Carcinoma Organoid Supplement B (50x)K2109-HN-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMHead and Neck Squamous Cell Carcinoma Organoid Supplement C (250x)K2109-HN-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Head and Neck Squamous Cell Carcinoma Organoid Complete MediumUse a sterile technique to prepare the head and neck squamous cell carcinoma organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1.Thaw Head and Neck Squamous Cell Carcinoma Organoid Supplement B(50x) and Head and Neck Squamous Cell Carcinoma Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL Head and Neck Squamous Cell Carcinoma Organoid Supplement B(50x) and 40μL Head and Neck Squamous Cell Carcinoma Organoid Supplement C(250x) to 9.76mL Head and Neck Squamous Cell Carcinoma Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The Head and Neck Squamous Cell Carcinoma Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived Head and Neck Squamous Cell Carcinoma OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human head and neck squamous cell carcinoma tissue pieces in ice-cold Primary Tissue Storage Solution(K601005)with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution(K601003)in a 15mL conical tube at 37°C, with variable incubation times ranging from 30 min to 1.5 h. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestion mixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010)to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well..CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 min to let the Matrigel solidify.12.Prepare the required amount of head and neck squamous cell carcinoma organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived head and neck squamous cell carcinoma organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥8 times every 2 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 7 min, as this may result in poororganoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm head and neck squamous cell carcinoma organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

浮游生物计数框 藻类定量由上海书培实验设备有限公司生产提供，规格有0.1ml 1ml 2ml 5ml 8ml ，用于蛔虫卵测定。介绍：配套产品有：不锈钢开口直壁容器产品名称：浮游生物计数框 藻类定量计数框规格：0.1ml 玻璃拼接框，0.1ml藻类不锈钢一体框，1ml 一体框，2ml 一体框，5ml一体框，S型/回型全玻璃计数框，S型/回型有机璃一体框材质：玻璃，有机玻璃规格：产品名称规格单价（元）品牌浮游生物计数框藻类定量计数框0.1ml 玻璃拼接框350上海书培0.1ml藻类不锈钢一体框400上海书培1ml 一体框350上海书培2ml 一体框350上海书培5ml一体框380上海书培S型/回型全玻璃计数框455上海书培S型/回型有机璃一体框550上海书培使用方法介绍：显微镜的校准：两重合线之间台尺格数目尺长度(mm)= - 两重合线之间目尺格数浮游生物计数时，要将样品充分摇匀，将样品置入计数框内，在显微镜或解剖镜下进行计数。用定量加样管在水样中部吸液移入计数框内。加样之前要将盖玻片斜盖在计数框上(如图)样品按准确定量注入，在计数框中一边进样，另一边出气，这样可避免气泡产生。注满后把盏玻片移正。(1)长条计数法首先将目测微尺放入目镜中，然后用台测微尺去校目尺的长度，再用S-R计数框计数，以目测微尺的长度作为“个长条的宽度，从计数框的左边直计数到计数框的右边称为一个长条。计数的长条数取决于浮游生物的多少，浮游生物越少，计数的长条就要越多，一般计数2~4个长条。计数时，浮游植物和浮游动物要分开计数，然后分别计算单位体积中的浮游植物数和浮游动物数。2）视野计数法先用台测微尺测出显微镜视野的直径，然后算出视野的面积，再用S-R计数框或网格计数框计数。计数时以视野为单位计数。1.其计算 公式为! Cx1000浮游生物个数/mL=I A-D-FA=。一个视野面积(mm2)D=视野的深度(mm) F=计数的视野数( -般至少10个)C=计数的生物个数。其计算公式为Cx1000浮游生物数/mL=LW-DSC=计数的浮游生物数L=一个长条的长度，也就是计数框的长度( m)W=一个长条的宽度，即目尺的长度(mm)D=一个长条的深度，即计数框的深度(m)S=计数的长条数。3,网格计数法如用网格计数框，可采用网格计数法。如浮游生物密度不大，可将框内生物全部数出，密度大时，可利用计数框上的刻度，计数其中的几行(如2.5.8行)其计算公式为：c.V1浮游生物数/升=C=计数的生物个数 V1=由1升水浓缩成的样品水量V2=计数的样品水量。小型:1沉降计数法：将水样或混合样取3个分样，分别装满3个等体积的沉降器中，加盖玻片静置24h后，使用倒置显微镜鉴定，计数。取样体积应视样品浊度和浮游植物密度而定。计算公式: C=N1VC-一单位体 积水体中标本总量，单位为个每毫升(cells/mL) :N三个 分样计数的标本总个数，单位为个(cells) V-一 三个分样的总体积，单位为毫升(mL)2浓缩计数法视样品中浮游植物数量多少，浓缩或稀释至适当体积，用取样管搅拌均匀，迅速将取样管直立于样品中，准确地1次吸取所需体积并移入浮游植物计数框，加盖玻片后进行鉴定、计数:浮游植物的计数视其数量多少确定计数全部、1/2或1/4, 重复计数4次.浮游动物每次的计数值应在100个以上。计算公式:网才样品C= (n*V1) 1 (V2*Vn)C-单位体积水体中标本总量， 单位为个每立方米(cells/m3) n一取样计数个数， 单位为个(cells) :V1-- -水样浓缩后的体积，单位为毫升(mL)↓V2-滤水量， 单位为立方米，Vn--取样计 数的体积，单位为毫升(mL) 。采水样品: C-单位体 积水体中标本总量，单位为个每升(cells/L) n-取样计数个数， 单位为个(cells) :V1-水样浓缩后的体积，单位为毫升(mL) V2-- 原采水量,单位为升(L) Vn-取样计 数的体积，单位为毫升(mL)。中型:1浮游动物体积分数测定优去除样品杂物，标定体积测量器的体积为50mL将样品全部倾入体积测量器内进行抽滤，样品中材网的水分滤出后，拧上底盖，再用装满50mL海水的滴定管从测量器的加水孔注入海水至液面与指针尖---端相接触为止。此时留在滴定管中的水量即代表浮游动物的体积，换算浮游动物体积分数(10 的负6次方)2浮游动物湿重含量测定去网孔略小于采样网孔的筛绢，剪成与漏斗内径相同的圆形，用水浸湿后沥干称重，记录。测定时，除去样品中杂物，将已标定重量的筛绢平铺于漏斗中，倒入样品抽滤片刻，移出载有样品的筛绢至吸 检器材水纸上吸去筛绢底表多余水分，然后称重。总重减去筛绢重即得样品湿重，换算浮游动物湿重生物含培界量(mg/m3 (立方米) ) 咨国公究3浮游动物干重含量测定方法基本同2,用已知重量的筛绢过滤样品，烘干后称重。总重减去筛绢重为浮游动物干重含量。换算含量(mg/m3)。4浮游生物个体计数：浮游生物计数框中，于体视显微镜下鉴定计数。

北京绿百草科技供应TOSOH正相/亲水色谱柱TSKGEL Amide-80HR，货号21982，4.6*250mm。TSKGEL Amide-80HR，粒径5&mu m，孔径100A。Amide-80HR以硅胶为基体，键合了氨基甲酰基的色谱柱，是对TSKgel Amide-80 5µ m进行了改良，提高了分离能力和耐久性，是一款高性能正相/亲水作用色谱柱。TSKGEL Amide-80主要分析糖类、多肽、核酸、亲水性药物等，日本药典中推荐使用TSKGEL Amide-80HR分离人参皂苷。北京绿百草科技可以提供Amide-80详细信息，还有相应的保护柱提供。

序号型号粒径规格备注18101（白色）40-60mesh60-80mesh80-100mesh25g硅藻土类19102（白色）40-60mesh60-80mesh80-100mesh25g硅藻土类206201（白色/红色）40-60mesh60-80mesh80-100mesh25g硅藻土类

水中尸体死因诊断是世界性的法医学难题，硅藻检验被公认为溺死诊断最可靠的方法（“金标准”）。传统硅藻检验方法因灵敏度低、易污染、劳动强度大、安全性差等原因而未能在法医学实践中得到广泛应用。我司研究的科技成果“微波消解-真空抽滤-扫描电镜联用硅藻检验方法”克服了传统方法的重大缺陷，具有灵敏、准确、安全、环保等优点，应用前景良好。硅藻检验样品盒是应用“微波消解-真空抽滤-扫描电镜联用硅藻检验方法”的重要工具，用于存放经微波消解、真空抽滤后得到的硅藻样品，具有操作简单、防损失、防污染、样品管理方便等特点，已获国家专利。硅藻检验样品盒不仅适用于法医学硅藻检验，亦可用于存放硅藻生态学研究、硅藻分类学研究等其它领域中的滤膜样品。硅藻检验样品盒采用一体化、紧凑式设计：一具尸体的检材（肺、肝、肾、骨髓、水样等）和控制样品经处理后所得到的样品，通常可集中存放于一个样品盒中。硅藻检验样品盒主要包含（由外到内）：1个环保型外盒：含密封性薄膜外包装、可供填写的样品信息栏、防揭标签等；1个内盒底座：含6个用于放置内盒的孔位；6个透明内盒；6个钉型扫描电镜样品座：样品座直径有25mm和28mm两种规格，上覆盖有EM级高纯碳导电胶。

水中尸体死因诊断是世界性的法医学难题，硅藻检验被公认为溺死诊断最可靠的方法（“金标准”）。传统硅藻检验方法因灵敏度低、易污染、劳动强度大、安全性差等原因而未能在法医学实践中得到广泛应用。我司研究的科技成果“微波消解-真空抽滤-扫描电镜联用硅藻检验方法”克服了传统方法的重大缺陷，具有灵敏、准确、安全、环保等优点，应用前景良好。硅藻检验样品盒是应用“微波消解-真空抽滤-扫描电镜联用硅藻检验方法”的重要工具，用于存放经微波消解、真空抽滤后得到的硅藻样品，具有操作简单、防损失、防污染、样品管理方便等特点，已获国家专利。硅藻检验样品盒不仅适用于法医学硅藻检验，亦可用于存放硅藻生态学研究、硅藻分类学研究等其它领域中的滤膜样品。硅藻检验样品盒采用一体化、紧凑式设计：一具尸体的检材（肺、肝、肾、骨髓、水样等）和控制样品经处理后所得到的样品，通常可集中存放于一个样品盒中。硅藻检验样品盒主要包含（由外到内）：1个环保型外盒：含密封性薄膜外包装、可供填写的样品信息栏、防揭标签等；1个内盒底座：含6个用于放置内盒的孔位；6个透明内盒；6个钉型扫描电镜样品座：样品座直径有25mm和28mm两种规格，上覆盖有EM级高纯碳导电胶。

北京绿百草科技供应TOSOH正相/亲水色谱柱 TSKGEL Amide-80HR，货号21982，4.6*250mm。TSKGEL Amide-80HR，粒径5&mu m，孔径100A。Amide-80HR以硅胶为基体，键合了氨基甲酰基的色谱柱，是对TSKgel Amide-80 5µ m进行了改良，提高了分离能力和耐久性，是一款高性能正相/亲水作用色谱柱。TSKGEL Amide-80主要分析糖类、多肽、核酸、亲水性药物等，日本药典中推荐使用TSKGEL Amide-80HR分离人参皂苷。北京绿百草科技可以提供Amide-80详细信息，还有相应的保护柱提供。货号柱名称官能团类型粒径尺寸mm21864Amide-80 (3)氨基甲酰 32.0*5021865Amide-80 (3)氨基甲酰 32.0*15021866Amide-80 (3)氨基甲酰 34.6*5021867Amide-80 (3)氨基甲酰 34.6*15020009Amide-80氨基甲酰 51.0*5020010Amide-80氨基甲酰 51.0*10021486Amide-80氨基甲酰 51.0*15021487Amide-80氨基甲酰 51.0*25019694Amide-80氨基甲酰 52.0*5019695Amide-80氨基甲酰 52.0*10019696Amide-80氨基甲酰 52.0*15019697Amide-80氨基甲酰 52.0*25013071Amide-80氨基甲酰 54.6*25021982Amide-80HR(5)氨基甲酰 54.6*25021967NH2-100 (3) 氨基 32.0*5021968NH2-100 (3) 32.0*15021969NH2-100 (3) 34.6*5021970NH2-100 (3) 34.6*15021862Guardgel Amide-80 (3), 2.0*10, 3pcs21863Guardgel Amide-80 (3), 3.2*15, 3pcs19308Guard Cartridge Holder - 2.0mmID19018Guard Cartridge Holder - 3.2mmID19010Guardgel Amide-80 (5), 3.2*15, 3pcs19021Guard Amide-80 4.6*1021941Guardgel Amide-80 (5), 2.0*10, 3pcs21971Guardgel NH2-100 (3), 2.0*10, 3pcs21972Guardgel NH2-100 (3), 3.2*15, 3pcs需要详细的信息请和绿百草科技联系：010-51659766登录网站获得更多产品信息:www.greenherbs.com.cn

Selector™ 环境类样品室与Selector™ 附件联用的环境类样品室配件是化学研究的理想选择特点：程序化控温最高达800°C压力范围从真空 (10-3Torr)到500 psi可控的压力环境(进气/出气)可靠的安全性能环境类样品室™ 此附件提供温度范围从室温到800°C，压力范围从真空(10-3Torr)到500 psi环境下的漫反射研究。标准样品室窗片ZnSe提供IR光信号传递及机械强度的良好平衡。可选配其他材质窗片。机身采用316不锈钢，提供机械强度及耐化学腐蚀性。低压能量源及过热自关闭系统保障安全性能。水冷舱保证高温操作下样品室外壁低温，放压系统保证高压状态下的安全性。与Selector™ 附件联用的环境类样品室配件是动力学、催化反应、表面分析、高分子材料及协调化学反应研究的理想选择。订购信息GS19930环境类样品室包含以下附件：高稳定性控制器及RS232界面请指定光谱仪制造厂家和型号。请指定220V或110V电压和使用的国家。可选配件GS28000 RS232连接包GS28001 USB连接包GS28002 RS485连接包耗材GS03610 KBr粉末(50g，光谱纯)GS19915 Selector™ 4mm 直径微型样品杯GS19916 Selector™ 11mm 直径标准样品杯GS19917带倾斜角度样品杯 -适用于总反射光收集GS19918 Selector™ 磨料样品盘架GS19919 Selector™ 12mm直径磨料样品盘（100）GS19931环境类样品室ESK

1ml网格定量藻类浮游生物计数框由上海书培实验设备有限公司生产提供，产品规格齐全，价廉物美，欢迎客户来电咨询选购。产品介绍：产品主要用于光学显微镜下鉴定和统计水样中的浮游植物（藻类）和小型浮游动物技术参数：产品名称：小型浮游生物计数框产品规格：1ml产品材质：进口优质玻璃框体：一体框网格数：常规款40格包装：1片/盒定做：可定做160格精细线（定制款详情联系我公司）

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍bioGenousTMOrganoid Dissociation Solution（类器官传代消化液）可应用于多种哺乳动物（如人、鼠、猪、蝙蝠、牛等）来源类器官的常规传代，可使类器官从基质胶中分离，并可使其温和地消化为小细胞簇或单细胞，同时保持其传代后的生长活力。技术参数产品信息产品组成货号规格储存条件及周期bioGenousTMOrganoid Dissociation SolutionE238001100mL2-8°C，12个月类器官消化过程中可能应用到的其他试剂试剂推荐货号Organoid Basal MediumB213152 & B213151Fetal Bovine Serum, FBS-类器官的消化传代1.向回收后的类器官中加入5-10倍类器官基质胶混合物体积的Organoid Dissociation Solution（类器官传代消化液），吹打混匀后在37°C条件下孵育1-8分钟使类器官解离（需提前取所需体积的消化液在37℃条件下预热，通常单层结构类器官的消化时间为0.5-3分钟，多层或者体积较大的类器官消化时间为3-8分钟）。注意：在此操作过程中须仔细监测消化过程，避免过度消化。在消化过程中，可使用移液器吹打混匀帮助消化。也可实时取少量消化悬液于显微镜下观察消化情况，当观察到较多的单细胞或直径在50μm以下的细胞簇后，即可认为消化完成。2.在确认消化完成的悬液中，加入至少五倍体积的类器官基础培养基进行稀释，从而终止消化作用。注意：时间较长的消化过程结束后可适当加入胎牛血清（Fetal Bovine Serum, FBS）至终浓度2%-5%以保证消化后细胞的活力。3.将上步骤所获类器官悬液进行离心（水平离心转子，150-300g，3分钟），弃上清，再次加入基础培养基重悬类器官沉淀。4.将上步骤所获类器官悬液进行离心（水平离心转子，150-300g，3分钟），弃上清后所获类器官可用于后续类器官培养、冻存等实验操作。

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。Product Description:bioGenousTMHepatocellular Carcinoma Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human hepatocellular carcinoma organoids. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMHepatocellular Carcinoma Organoid Basal MediumK2105-HCC-A100/A500100mL/500mL4℃，12 monthsbioGenousTMHepatocellular Carcinoma Organoid Supplement B (50x)K2105-HCC-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMHepatocellular Carcinoma Organoid Supplement C (250x)K2105-HCC-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Hepatocellular Carcinoma Organoid Complete MediumUse a sterile technique to prepare the hepatocellular carcinoma organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1.Thaw Hepatocellular Carcinoma Organoid Supplement B(50x) and Hepatocellular Carcinoma Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL Hepatocellular Carcinoma Organoid Supplement B(50x) and 40μL Hepatocellular Carcinoma Organoid Supplement C(250x) to 9.76mL Hepatocellular Carcinoma Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The Hepatocellular Carcinoma Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived Hepatocellular Carcinoma OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human hepatocellular carcinoma tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution (K601003) in a 15mL conical tube at 37°C, with variable incubation times ranging from 15 min to 45 min. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestionmixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010) to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well..CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 minto let the Matrigel solidify.12.Prepare the required amount of hepatocellular carcinoma organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived hepatocellular carcinoma organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥5 times every 1 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 3 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm hepatocellular carcinoma organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.25.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMCholangiocarcinoma Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human cholangiocarcinoma organoids. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMCholangiocarcinoma Organoid Basal MediumK2104-LB-A100/A500100mL/500mL4℃，12 monthsbioGenousTMCholangiocarcinoma Organoid Supplement B (50x)K2104-LB-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMCholangiocarcinoma Organoid Supplement C (250x)K2104-LB-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Cholangiocarcinoma Organoid Complete MediumUse a sterile technique to prepare the cholangiocarcinoma organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1. Thaw Cholangiocarcinoma Organoid Supplement B(50x) and Cholangiocarcinoma Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL Cholangiocarcinoma Organoid Supplement B(50x) and 40μL Cholangiocarcinoma Organoid Supplement C(250x) to 9.76mL Cholangiocarcinoma Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The Cholangiocarcinoma Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived Cholangiocarcinoma OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human cholangiocarcinoma tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution (K601003) in a 15mL conical tube at 37°C, with variable incubation times ranging from 30 min to 1.5 h. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestion mixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010) to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well..CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 min to let the Matrigel solidify.12.Prepare the required amount of cholangiocarcinoma organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived cholangiocarcinoma organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥8 times every 2 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 5 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm cholangiocarcinoma organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

产品特点：干基质点样Bond Elut 干基质点样（DmS）近年来干血斑技术在DMPK/ADME 中的应用已经获得重大突破。样品采集、运输和储存的实用性优势为大型药学实验室和CRO 实验室提供了显著的资源优势。Bond Elut DMS 为干血斑技术的应用重新设定了性能标准。创新的非纤维素点样材料提供了关键的灵敏度和工作流程上的优势，弥补了现有纤维素血液点样卡的主要不足。* 非纤维素“点样卡”减少了非特异性结合，改善了MS 分析响应，提高了信噪比* 简便的产品选择——针对多种生物基质样品处理的快速方法开发，仅需单一的、未经处理的点样卡* 宽范围的血细胞比容水平条件下，点样尺寸、均匀性和回收率具有很高的重现性：您可以满怀信心地针对多种生物基质样品进行分析方法开发* 非吸湿性材料即使在极湿的环境中也不会吸潮，减小了样品在运输/储存中存在的潜在危险，保证了其稳定性* 该DMS 卡需要的打孔力度仅为纤维素类DBS 卡的五分之一，确保更轻松的工作流程和高度的自动化兼容性干基质点（DMS）样品前处理卡50/包 A400150订购信息：Bond Elut 干基质点样（DMS）说明单位部件号干基质点（DMS）样品前处理卡50/包A400150干基质点（DMS）样品前处理卡500/包A400150KDMS 附件包包括 5 x 3 mm 打孔工具和 5 个打孔垫A42001 DMS 启始工具包包括 1 个带 5 x 3 mm 打孔工具和 5 个打孔垫的附件包（部件号：A42001）和 Bond Elut DMS 卡，50 个/包（部件号：A400150）A400150SK

样本台（样本座，specimen mount）样品大小要适合仪器专用样品台的尺寸，不能过大，样本座（样本台）尺寸各仪器均不相同，一般小的样品座为Φ3～5mm，大的样品座为Φ30～50mm，以分别用来放置不同大小的样本，样本的高度也有一定的限制，一般在5～10mm左右。扫描电镜的块状样品制备较简单，对于块状导电材料，除了大小要适合仪器样品座尺寸外，基本上不需进行什么制备，用导电胶把样品粘结在样品座上， 即可放在扫描电镜中观察。对于块状的非导电或导电性较差的材料，要先进行镀膜处理，在材料表面形成一层导电膜，以避免电荷积累，影响图象质量，并可防止样品的热损伤。粉末样本的制备：先将导电胶或双面胶纸粘结在样品座上，再均匀地把粉末样撒在上面，用洗耳球吹去未粘住的粉末，再镀上一层导电膜，即可上电镜观察。海德提供全系列的样品座（样本台）产品供大家选择！客户购买样本台，首先要知道所用仪器的生产厂家，仪器型号，样本台上下面的直径、材质等。我公司经销的样品台可以匹配的仪器生厂家或者仪器型号有：l AMRAY 1000/1200l AMRAY 1000/1200/1400l Cambridge S-4, Mark II, S-410...l Cambridge, Phillips, Camscan, B&L, Etec, Zeiss, etc.l Leo Microscopel Coates & Welterl Hitachi S-450l Hitachil Hitachi S-500l Jeoll Jeol, ISIl Jeol JSM 840l Zeiss产品信息1. Flat End Pin，Surface: ½" dia. (12.7mm) Slotted Head, 1/8" dia. (3.1mm) 货号产品名称规格75500Specimen Mount50/pk75510Specimen Mount100/pk75520Specimen Mount500/pk2.Aluminum slotted head. Surface 1" (25.4mm) x 1⁄2" (12.7mm) H, pin 1⁄8" (3.1mm) Dia ，适于以下品牌Leica, Cambridge, LEO, FEI, Philips, Zeiss, Camscan货号产品名称规格75183Specimen Mount10/pk75184Specimen Mount50/pk75185Specimen Mount100/pk3.Aluminum large slotted head. Surface 1 1⁄4" (32mm) x 1⁄2" 12.7mm) H, pin 1⁄8" (3.1mm) Dia。适于以下品牌Leica, Cambridge, LEO, FEI, Philips, Zeiss, Camscan货号产品名称规格75187Specimen Mount10/pk75188Specimen Mount50/pk75189Specimen Mount100/pk

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMBreast Cancer Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human breast cancer organoids. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMBreast Cancer Organoid Basal MediumK2147-BC-A100/A500100mL/500mL4℃，12 monthsbioGenousTMBreast Cancer Organoid Supplement B (50x)K2147-BC-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMBreast Cancer Organoid Supplement C (250x)K2147-BC-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Breast Cancer Organoid Complete MediumUse a sterile technique to prepare the breast cancer organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1.Thaw Breast Cancer Organoid Supplement B(50x) and Breast Cancer Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL Breast Cancer Organoid Supplement B(50x) and 40μL Breast Cancer Organoid Supplement C(250x) to 9.76mL Breast Cancer Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The Breast Cancer Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived Breast Cancer OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human breast cancer tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution (K601003) in a 15mL conical tube at 37°C, with variable incubation times ranging from 30 min to 1.5 h. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestionmixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010) to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well.CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 minto let the Matrigel solidify.12.Prepare the required amount of breast cancer organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived breast cancer organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥8 times every 2 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 7 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm breast cancer organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.25.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMOvarian Cancer Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human Ovarian Cancer organoids. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMOvarian Cancer Organoid Basal MediumK2168-OC-A100/A500100mL/500mL4℃，12 monthsbioGenousTMOvarian Cancer Organoid Supplement B (50x)K2168-OC-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMOvarian Cancer Organoid Supplement C (250x)K2168-OC-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Ovarian Cancer Organoid Complete MediumUse a sterile technique to prepare the ovarian cancer organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1.Thaw Ovarian Cancer Organoid Supplement B(50x) and Ovarian Cancer Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL Ovarian Cancer Organoid Supplement B(50x) and 40μL Ovarian Cancer Organoid Supplement C(250x) to 9.76mL Ovarian Cancer Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The Ovarian Cancer Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived Ovarian Cancer OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human ovarian cancer tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution (K601003) in a 15mL conical tube at 37°C, with variable incubation times ranging from 30 min to 1.5 h. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestionmixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010) to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well..CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 minto let the Matrigel solidify.12.Prepare the required amount of ovarian cancer organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived ovarian cancer organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥8 times every 2 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 7 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm ovarian cancer organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.25.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMGastric Cancer Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human gastric cancer organoid s. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMGastric Cancer Organoid Basal MediumK2179-GC-A100/A500100mL/500mL4℃，12 monthsbioGenousTMGastric Cancer Organoid Supplement B (50x)K2179-GC-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMGastric Cancer Organoid Supplement C (250x)K2179-GC-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Gastric Cancer Organoid Complete MediumUse a sterile technique to prepare the gastric cancer organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1.Thaw Gastric Cancer Organoid Supplement B(50x) and Gastric Cancer Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL Gastric Cancer Organoid Supplement B(50x) and 40μL Gastric Cancer Organoid Supplement C(250x) to 9.76mL Gastric Cancer Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The Gastric Cancer Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived Gastric Cancer OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human gastric cancer tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution (K601003) in a 15mL conical tube at 37°C, with variable incubation times ranging from 30 min to 1 h. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestionmixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010) to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well..CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 minto let the Matrigel solidify.12.Prepare the required amount of gastric cancer organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived gastric cancer organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥8 times every 2 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 5 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm gastric cancer organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.25.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMHuman Liver Ductal Organoid Kit is a chemically defined cell culture medium for establishment and maintenance of human liver ductal organoids(hLDs) derived from adult stem cells. Self-renewal of the liver ductal epithelium is driven by the proliferation of stem cells and their progenitors located in liver. hLDs display all hallmarks of the liver ductal epithelium in terms of architecture, cell type composition, and self-renewal dynamics, therefore hold great promise for unprecedented studies of Human liver development and disease, hLDs could differentiate into hepatocyte like cell under the induction of differentiation medium.技术参数Product Information:ComponentComponent Cat#VolumeStorage& StabilitybioGenousTMHuman Liver Ductal Organoid Basal Medium（Differentiation）K2008-HLH –A100/A500100mL/500mL4℃，12 monthsbioGenousTMHuman Liver Ductal Organoid Supplement B(50x)（Differentiation）K2008-HLH –B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMHuman Liver Ductal Organoid Supplement C(250x)（Differentiation）K2008-HLH –C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMHuman Liver Ductal Organoid Supplement D(250x)（Differentiation）K2008-HLH –D100/D5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Human Liver Ductal Organoid Differentiation MediumUse sterile technique to prepare the human liver ductal organoid differentiation medium. hLDs grown in Human Liver Ductal Organoid Expansion Medium overwhelmingly consisted of cholangiocytes. After changing the Expansion Medium to differentiation medium, the hLDs could differentiate into hepatocyte like cells, which display the markers of hepatocytes, includingALBUMIN,TTRandCYP3A4. The following examples are for preparing 10 mL of Differentiation I Medium and Differentiation II Medium. If preparing other volumes, adjust accordingly.1.Thaw Human Liver Ductal Organoid Supplement B(50x) (Differentiation), Human Liver Ductal Organoid Supplement C(250x) (Differentiation) and Human Liver Ductal Organoid Supplement D(250x) (Differentiation) on ice.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.For Human Liver Ductal Organoid Differentiation Medium I. Add 40 μL Human Liver Ductal Organoid Supplement D(250x) (Differentiation) to 10 mL Human Liver Ductal Organoid Medium(Expansion) (K2008-HLD). Mix thoroughly.3.For Human Liver Ductal Organoid Differentiation Medium II. Add 200 μL Human Liver Ductal Organoid Supplement B(50x) (Differentiation) and 40 μL Human Liver Ductal Organoid Supplement C(250x)（Differentiation）to 10 mL Human Liver Ductal Organoid Basal Medium (Differentiation). Mix thoroughly.NOTE:If not use immediately, store complete medium at 2-8°C for not more than 2 weeks. bioGenousTMHuman Liver Ductal Organoid Supplement B (Differentiation) contains fungicide and antibiotics(50x).

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍bioGenousTMOrganoid Cryopreservation Medium（类器官冻存液）可应用于多种哺乳动物（如人、鼠、猪、蝙蝠、牛等）来源的类器官和细胞系的低温长期保存，该冻存液不含血清，不含任何动物来源成分，含有10% DMSO。技术参数产品信息产品组成货号规格储存条件及周期bioGenousTMOrganoid Cryopreservation Medium（Serum Free）E238023100mL4℃，3年类器官（或细胞）的冻存和复苏需选用生长状态良好的类器官（或细胞）进行冻存实验。类器官（或细胞）的冻存1.向回收后的类器官（或细胞）中加入预冷的（2-8℃）Organoid Cryopreservation Medium（类器官冻存液），吹打混匀后迅速转移至细胞冻存管中（冻存管内细胞悬液体积应大于0.5mL）。注意：每1mL冻存液推荐冻存1x103-1x107个细胞或对应细胞量的类器官。2.将冻存管放入细胞冻存程序降温盒内（降温盒须提前平衡至室温或4℃），随后立即将降温盒放入-80 ℃超低温冰箱中；也可将冻存管进行人工梯度降温处理，如4℃静置10分钟，-20℃保存1小时，-80℃保存过夜。3.次日或12小时后将冻存管于低温条件下（不高于-70℃，推荐使用干冰或原降温盒）转移至液氮中长期保存。类器官（或细胞）的复苏4.提前在37℃条件下预热类器官（或细胞）所需的基础培养基。5.在37℃的水浴中快速解冻细胞冻存管，当冻存管内冻存物仅剩些许冰渣残留时立即停止水浴并及时转移至洁净操作台。6.将冻存悬液转移至离心管中，缓缓加入5-10倍体积的预热的基础培养基，轻轻混匀。7.将上步骤所获类器官（或细胞）悬液进行离心（水平离心转子，150-300g，3分钟），弃上清，再次加入基础培养基重悬类器官（或细胞）沉淀。8.将上步骤所获类器官（或细胞）悬液进行离心（水平离心转子，150-300g，3分钟），弃上清后所获类器官（或细胞）可用于后续类器官（或细胞）的培养。

抓取杆的螺丝可以拧入160-41圆形冷冻样品盒的螺纹中心，用于移动样品盒。可用于将冷冻样品盒存储在液氮中。产品编号描述单位160-46冷冻样品盒抓取杆，白色个160-46-4冷冻样品盒抓取杆，黄色个160-46-6冷冻样品盒抓取杆，蓝色个160-46-0冷冻样品盒抓取杆，黑色个

产品名称：Semi-demountable Torches仪器厂商：Agilent/美国 安捷伦价格：面议库存：是DescriptionApplicationPart No.Semi-demountable inertaxial torch assembly kitSamples with free HF when used with an inertalumina injector (2410067590) for volatile organicsolvents, use narrow bore 0.8 mm ID quartz injector(2010077500)9910084700Semi-demountable inertradial torch assembly kitSamples with free HF when used with an inertalumina injector (2410067590) for volatile organicsolvents, use narrow bore 0.8 mm ID quartz injector(2010077500)9910056800

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMSmall Cell Lung Cancer Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human Small Cell Lung Cancer organoids. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMSmall Cell Lung Cancer Organoid Basal MediumK2121-SL-A100/A500100mL/500mL4℃，12 monthsbioGenousTMSmall Cell Lung Cancer Organoid Supplement B (50x)K2121-SL-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMSmall Cell Lung Cancer Organoid Supplement C (250x)K2121-SL-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Small Cell Lung Cancer Organoid Complete MediumUse a sterile technique to prepare the small cell lung cancer organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1.Thaw Small Cell Lung Cancer Organoid Supplement B(50x) and Small Cell Lung Cancer Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL Small Cell Lung Cancer Organoid Supplement B(50x) and 40μL Small Cell Lung Cancer Organoid Supplement C(250x) to 9.76mL Small Cell Lung Cancer Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The Small Cell Lung Cancer Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived Small Cell Lung Cancer OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human small cell lung cancer tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution (K601003) in a 15mL conical tube at 37°C, with variable incubation times ranging from 30 min to 1 h. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestionmixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010) to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well..CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 minto let the Matrigel solidify.12.Prepare the required amount of small cell lung cancer organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived small cell lung cancer organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥3 times every 1 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 3 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm small cell lung cancer organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.25.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMEndometrialCancer Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human endometrialcancer organoids. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMEndometrialCancer Organoid Basal MediumK2166-EC-A100/A500100mL/500mL4℃，12 monthsbioGenousTMEndometrialCancer Organoid Supplement B (50x)K2166-EC-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMEndometrialCancer Organoid Supplement C (250x)K2166-EC-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of EndometrialCancer Organoid Complete MediumUse a sterile technique to prepare the endometrialcancer organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1.Thaw EndometrialCancer Organoid Supplement B(50x) and EndometrialCancer Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL EndometrialCancer Organoid Supplement B(50x) and 40μL EndometrialCancer Organoid Supplement C(250x) to 9.76mL EndometrialCancer Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The EndometrialCancer Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived EndometrialCancer OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human endometrialcancer tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution (K601003) in a 15mL conical tube at 37°C, with variable incubation times ranging from 30 min to 1 h. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestion mixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010) to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well..CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 min to let the Matrigel solidify.12.Prepare the required amount of endometrialcancer organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived endometrialcancer organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥8 times every 2 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 7 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant andresuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm endometrialcancer organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。Product Description:bioGenousTMLung Adenocarcinoma Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human lung adenocarcinoma organoids. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMLung Adenocarcinoma Organoid Basal MediumK2138-LA-A100/A500100mL/500mL4℃，12 monthsbioGenousTMLung Adenocarcinoma Organoid Supplement B (50x)K2138-LA-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMLung Adenocarcinoma Organoid Supplement C (250x)K2138-LA-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Lung Adenocarcinoma Organoid Complete MediumUse a sterile technique to prepare the lung adenocarcinoma organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1.Thaw Lung Adenocarcinoma Organoid Supplement B(50x) and Lung Adenocarcinoma Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL Lung Adenocarcinoma Organoid Supplement B(50x) and 40μL Lung Adenocarcinoma Organoid Supplement C(250x) to 9.76mL Lung Adenocarcinoma Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The Lung Adenocarcinoma Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived Lung Adenocarcinoma OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human lung adenocarcinoma tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution (K601003) in a 15mL conical tube at 37°C, with variable incubation times ranging from 30 min to 1.5 h. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestion mixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010) to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well..CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 min to let the Matrigel solidify.12.Prepare the required amount of lung adenocarcinoma organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived lung adenocarcinoma organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥8 times every 2 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 7 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm lung adenocarcinoma organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.25.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.

类器官（Organoids）是指将成体干细胞或多能干细胞在体外三维培养形成的具有一定空间结构的组织类似物。类器官在组织结构、细胞类型、自我更新能力和功能等方面与来源组织高度一致，从而在发育生物学、疾病造模、精准医学、药物研发、基因和细胞疗法、感染和免疫以及再生医学等生物医学的多个领域展现出独特的优势。产品介绍Product Description:bioGenousTMCervical Cancer Organoid Kit is a chemically defined cell culture medium for the establishment and maintenance of human cervical cancer organoids. Patient-derived cancer organoids recapitulate the genomic and pathological features of original tumors and therefore hold great promise for medical research and precision medicine.技术参数Product Information:ComponentComponent Cat#VolumeStorage & StabilitybioGenousTMCervical Cancer Organoid Basal MediumK2169-CC-A100/A500100mL/500mL4℃，12 monthsbioGenousTMCervical Cancer Organoid Supplement B (50x)K2169-CC-B100/B5002mL/10mL-20℃,avoid repeated freeze-thaw cycles, 12 monthsbioGenousTMCervical Cancer Organoid Supplement C (250x)K2169-CC-C100/C5000.4mL/2mL-20℃, avoid repeated freeze-thaw cycles, 12 monthsPreparation of Cervical Cancer Organoid Complete MediumUse a sterile technique to prepare the cervical cancer organoid complete medium. The following example is for preparing 10mL complete medium. If preparing other volumes, adjust accordingly.1.Thaw Cervical Cancer Organoid Supplement B(50x) and Cervical Cancer Organoid Supplement C(250x) on ice. Mix thoroughly.NOTE:Once thawed, use immediately or aliquot and store at -20°C for not more than 10 months. After thawing the aliquots, use immediately. Do not re-freeze.2.Add 200μL Cervical Cancer Organoid Supplement B(50x) and 40μL Cervical Cancer Organoid Supplement C(250x) to 9.76mL Cervical Cancer Organoid Basal Medium. Mix thoroughly.NOTE:If not used immediately, store the complete medium at 2-8°C for not more than 2 weeks. The Cervical Cancer Organoid Supplement B contains fungicide and antibiotics (50x).Protocol for Establishment ofPatient-Derived Cervical Cancer OrganoidsCAUTIONStudies involving primary human tissue material must follow all relevant institutional and government regulations. Informed consent must be obtained from all subjects before the collection of the primary human tissue material.Establishment of Organoids from Primary Tissue1.Collect primary human cervical cancer tissue pieces in ice-cold Primary Tissue Storage Solution (K601005) with conical tubes. Keep tissue samples at 4°C until the start of the isolation.2.Assess whether the obtained tissue pieces consist purely of epithelium. If fat or muscle tissues are present, remove these non-epithelial components as much as possible using surgical scissors or scalpels and forceps under a dissection microscope. If no fat or muscle tissues are present, continue to the next step immediately.3.Rinse the tissuewith Cancer Organoid Basal Medium (B213152) or DPBS twice.4.Mince the tissue into small fragments of 1-3 mm3in a cell culture dish using surgical scissors or scalpels.5.Digest the tissue fragments with 10mL of Tumor Tissue Digestion Solution (K601003) in a 15mL conical tube at 37°C, with variable incubation times ranging from 30 min to 1 h. Carefully monitor the digestion process, mixing the content of the tube every 5-10 min by shaking vigorously or pipetting the mixture up and down. The digestion process could be finished when most of tissue fragments are able to pass through the 1mL pipette tips.6.Add FBS to the tissuedigestionmixture to a final concentration of 2%, and filter using a 100 μm cell strainer.7.Collect and centrifuge the filtered cells at 250g for 3 min at 4 °C. In case of a visible red pellet, aspirate the supernatant, and resuspend the pellet using 2mL of Red Blood Cell Lysis Solution (E238010) to lyse the erythrocytes at room temperature for 1 min and centrifuge at 250g for 3 min at 4°C.8.Aspirate the supernatant and resuspend the pellet in Cancer Organoid Basal Medium, centrifuge at 250g for 3 min at 4°C,repeat this step once more time.9.Aspirate the supernatant and resuspend the pellet in Matrigel. The Matrigel should be kept on ice to prevent it from solidifying. Perform the process as quickly as possible. The amount of Matrigel used depends on the size of the pellet. Approximately 10,000 cells should be plated in 25 μL of Matrigel.CRITICAL:Do not overly dilute the Matrigel (Matrigel should be 70% (Matrigel vol/Total vol)), as this may inhibit the proper formation of the solid droplets.10.Plate the Matrigel containing organoids at the bottom of 24-well cell culture plates in droplets of ~30μL each around the center of the well..CRITICAL:Once the organoids are resuspended in Matrigel, proceed with plating as quickly as possible, as the Matrigel may solidify in the tube or pipette tip. Do not let the Matrigel touch the wall of wells.11.Place the culture plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for15-25 minto let the Matrigel solidify.12.Prepare the required amount of cervical cancer organoid complete medium.13.Once the Matrigel droplets have solidified (15-25 min), open the plate and carefully add 500 μL of organoid complete medium to each well.CRITICAL:Do not add the medium directly on top of the Matrigel droplets, as this might disrupt the droplets.14.Place the culture plate in a humidified incubator at 37 °C and 5% (vol/vol) CO2.15.Change the medium every 3-4 d by carefully aspirating the medium from the wells and replacing it with a fresh, pre-warmed organoid complete medium.16.Closely monitor the organoid formation. Ideally, patient-derived cervical cancer organoids should be passaged for the first time between 7 and 10 d after the initial plating.Splitting and Passaging of Organoids17.Pipette up and down to disrupt the Matrigel and transfer the organoid suspension to a 1.5 mL conical tube.18.Pipette the organoid suspension up and down to mix thoroughly by pipetting against the bottom of the tube to create pressure, which will aid the removal of Matrigel.19.Centrifuge organoids at 250g for 3 min at room temperature.20.Aspirate the supernatant and split organoids using either Organoid Dissociation Solution (E238001) or by mechanical disruption. For Organoid Dissociation Solution-based cell dissociation, resuspend the pellet in 0.2 mL of Organoid Dissociation Solution, pipette up and down and incubate at 37 °C until organoids fall apart. Pipette up and down with a filter tip for ≥8 times every 2 min to aid in the disruption of the organoids. Closely monitor the digestion to keep the incubation time inthe Organoid Dissociation Solution to a minimum. In case of mechanical disruption, resuspend the pellet in 1.5 mL ofCancer Organoid Basal Medium. Carefully pipette the organoid suspension up and down 30 times by pipetting against the bottom of the tube to create pressure, which will aid organoid disruption.CRITICAL:Do not dissociate in Organoid Dissociation Solution for 7 min, as this may result in poor organoid outgrowth or even loss of the culture. As a rule of thumb, digestion is complete if a mixture of small lumps of cells (consisting of 10–50 cells) can be observed.21.After shearing is complete, wash once by topping up with 1 mL ofCancer Organoid Basal Medium, and centrifuge at 250g for 3 min at room temperature.22.Aspirate the supernatant and resuspend the organoid pellet in 70% (vol/vol) Matrigel, and plate organoids in droplets at the bottom of a culture plate as described in Steps 10. After plating is complete, transfer the plate into a humidified incubator at 37 °C and 5% (vol/vol) CO2for 15–25 min.23.Pre-warm cervical cancer organoid complete medium at 37 °C.24.After the Matrigel droplets have solidified (15–25 min), carefully pipette pre-warmed medium into the wells.25.Place culture plates in a humidified incubator at 37 °C and 5% (vol/vol) CO2until the organoids are needed for further experiments.