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Keratinocyte Growth Kit
PCS-200-040™
Keratinocyte Growth Kit (ATCC PCS-200-040) contains components that when added to Dermal Cell Basal Medium (ATCC PCS-200-030) create a complete ATCC® Primary Cell Solutions™ culture environment for keratinocytes derived from normal human epidermis (e.g., Primary Epidermal Keratinocytes, Normal, Human Neonatal Foreskin, ATCC PCS-200-010). The complete serum-free (although not animalfree) media is formulated to inhibit fibroblast growth, and the low calcium concentration (0.06 mM) slows differentiation. No feeder layers, extracellular matrix proteins or other substrates are required. The only animal-derived component is Extract P; if necessary, keratinocytes can be cultured in the absence of Extract P.
Product category
Reagents
Comments
The Keratinocyte Growth Kit contains components that when added to Dermal Cell Basal Medium (ATCC PCS-200-030) create a complete serum-free culture environment for keratinocytes. The complete keratinocyte growth medium is formulated to inhibit fibroblast growth, and the low calcium concentration (0.06 mM) slows differentiation. No feeder layers, extracellular matrix proteins or other substrates are required. The final concentration for each component in complete keratinocyte growth medium is as follows:
Extract P: 0.4%
rh TGF-alpha: 0.5 ng/mL
L-Glutamine: 6 mM
Hydrocortisone: 100 ng/mL
Insulin: 5 µg/mL
Epinephrine: 1.0 µM
Apo-transferrin: 5 µg/mL
Applications
Cell growth and viability
Product format
Frozen
Shipping information
1 kit
Storage conditions
-20°C or colder, -70°C for long-term storage
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Documentation
Product sheet
Certificate of analysis
Certificate of origin
Safety data sheet
Biosafety Icon BSL 1
ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.
Handling procedure
Unpacking and Storage Instructions
Check all containers for leakage or breakage.
Store the growth kit(s) at either -20°C in a freezer that is not self-defrosting or at -70°C for long-term storage. If thawed upon arrival, the Growth Kit can be stored at 2°C to 8°C and added to the basal medium within 72 hours of receipt.
Preparation of Complete Growth Media
Obtain one growth kit from the freezer; make sure that the caps of all components are tight.
Thaw the components of the growth kit just prior to adding to the basal medium. It is necessary to warm the L-glutamine component in a 37°C water bath and shake to dissolve any precipitates, prior to adding to the basal medium.
Obtain one bottle of Dermal Cell Basal Medium (485 mL) from cold storage.
Decontaminate the external surfaces of all growth kit component vials and the basal medium bottle by spraying with 70% ethanol.
Using aseptic technique and working in a laminar flow hood or biosafety cabinet, transfer the indicated volume of each growth kit component to the bottle of basal medium using a separate sterile pipette for each.
Tightly cap the bottle of complete growth medium and swirl the contents gently to assure a homogeneous solution. Do not shake forcefully to avoid foaming. Label and date the bottle.
Complete media should be stored in the dark at 2°C to 8°C (do not freeze). When stored under these conditions, complete media is stable for 30 days.
Table 1. Keratinocyte Growth Kit Components
Component
Volume
Final Concentration
Extract P
2.0 mL
0.4%
rh TGF-a
0.5 mL
0.5 ng/mL
L-Glutamine
15.0 mL
6 mM
Hydrocortisone Hemisuccinate
0.5 mL
100 ng/mL
rh Insulin
0.5 mL
5 µg/mL
Epinephrine
0.5 mL
1.0 µM
Apo-Transferrin
0.5 mL
5 µg/mL
Antimicrobials and phenol red are not required for proliferation, but may be added if desired. The recommended volume of each optional component to be added to the complete growth media is summarized in Table 2.
Table 2. Addition of Antimicrobials/Antimycotics and Phenol Red (Optional)
Component
Volume
Final Concentration
Gentamicin-Amphotericin B Solution
0.5 mL
Gentamicin: 10 µg/mL
Amphotericin B: 0.25 µg/mL
Penicillin-Streptomycin-Amphotericin B Solution
0.5 mL
Penicillin: 10 Units/mL
Streptomycin: 10 µg/mL
Amphotericin B: 25 ng/mL
Phenol Red
0.5 mL
33 µM
Cryopreservation
1. Check all containers for leakage or breakage.
2. Store the growth kit(s) at either -20°C in a freezer that is not self-defrosting or at -70°C for long-term storage. If thawed upon arrival, the Growth Kit can be stored at 2°C to 8°C and added to the basal medium within 72 hours of receipt.
Quality control specifications
Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Functional tests
Rate of proliferation and morphology
Legal disclaimers
Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty
The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.
Disclaimers
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.
While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.
This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.
Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.
