Dermal Cell Basal Medium

PCS-200-030 ^™

Dermal Cell Basal Medium is a sterile, phenol-red-free, liquid tissue culture medium intended for use as one component in a complete ATCC Primary Cell Solutions system. This system is designed to support cells (eg., keratinocytes, melanocytes) derived from a normal human epidermis. Dermal Cell Basal Medium contains essential and non-essential amino acids, vitamins, other organic compounds, trace minerals, and inorganic salts.

Product category: Media
Applications: Cell growth and viability
Volume: 485 mL
Storage conditions: 2°C to 8°C

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Documentation

Product sheet

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.

General

Specific applications

Dermal Cell Basal Medium is a sterile, phenol red-free, liquid tissue culture medium intended for use as one component in a complete ATCC Primary Cell Solutions™ system.

This system is designed to support cells (e.g., keratinocytes, melanocytes) derived from normal human epidermis. Dermal Cell Basal Medium contains essential and non-essential amino acids, vitamins, other organic compounds, trace minerals and inorganic salts.

To support the proliferation and plating efficiency of keratinocytes (e.g., Primary Epidermal Keratinocytes, Normal, Human Neonatal Foreskin, ATCC PCS-200-010), Dermal Cell Basal Medium must be supplemented with the Keratinocyte Growth Kit (ATCC PCS-200-040). The complete serum-free (although not animal-free) media is formulated to inhibit fibroblast growth, and the low calcium concentration (0.06 mM) slows differentiation.

To support the proliferation and plating efficiency of melanocytes (e.g., Primary Epidermal Melanocytes, Normal, Human Neonatal, ATCC PCS-200-012 or Primary Epidermal Melanocytes, Normal, Human Adult, ATCC PCS-200-013), Dermal Cell Basal Medium must be supplemented with either the Melanocyte Growth Kit (ATCC PCS-200-041) or the Adult Melanocyte Growth Kit (ATCC PCS-200-042).

Using Dermal Cell Basal Medium supplemented with the appropriate growth kit, the growth of keratinocytes and melanocytes are supported without the use of feeder layers, extracellular matrix proteins or other substrates.

Optional media supplements:

Gentamicin-Amphotericin B Solution (ATCC PCS-999-025)
Penicillin-Streptomycin-Amphotericin B Solution (ATCC PCS-999-002)
Phenol Red (ATCC PCS-999-001)

Handling information

Handling procedure

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 them 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 them with 70% ethanol.
Using aseptic technique and working in a laminar flow hood or biosafety cabinet, transfer the volume of each growth kit component, as indicated in Table 1 or 2, to the bottle of basal medium using a separate sterile pipette for each transfer

Table 1. If using the Keratinocyte Growth Kit (ATCC^® PCS-200-040), add the indicated volume for each component:

Component	Volume	Final Concentration
Bovine Pituitary Extract (BPE)	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

Table 2. If using the Melanocyte Growth Kit (ATCC^® PCS-200-041), add the indicated volume for each component:

Component	Volume	Final Concentration
rh Insulin	0.5 mL	5 mg/mL
Ascorbic Acid	0.5 mL	50 mg/mL
L-Glutamine	15.0 mL	6 mM
Epinephrine	0.5 mL	1.0 mM
Calcium chloride	80 mL	0.2 mM
M8 Supplement	5 mL	Proprietary formulation

Table 3. If using the Adult Melanocyte Growth Kit (ATCC^® PCS-200-042), add the indicated volume for each component:

Component	Volume	Final Concentration
rh Insulin	0.5 mL	5 mg/mL
Ascorbic Acid	0.5 mL	50 mg/mL
L-Glutamine	15.0 mL	6 mM
Epinephrine	0.5 mL	1.0 mM
Calcium Chloride	840 mL	1.5 mM
Peptide Growth Factor	1.0 mL	Proprietary formulation
M8 Supplement	5 mL	Proprietary formulation

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 media is summarized in Table 4.

Table 4. Addition of Antimicrobials/Antibiotics 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

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 growth 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.

Cryopreservation

1. Check all containers for leakage or breakage.

2. Store the basal medium at 2°C to 8°C. Do not freeze. Protect from light. Do not exceed 37°C when warming the media prior to use. When stored in the dark at 2°C to 8°C, the product is stable until the expiration date on the label.

Quality control specifications

Bacterial and fungal testing: Not detected
Mycoplasma contamination: Not detected
Endotoxin: < 0.5 EU/mL
Osmolality: 315 ± 10 mOsm/kg
pH: 7.5 ± 0.2
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.

References

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