Adult Melanocyte Growth Kit

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.

Preparation of Complete Growth Media

1. Obtain one growth kit from the freezer; make sure that the caps of all components are tight.
2. Thaw the components of the growth kit just prior to adding them to the basal medium.
3. Obtain one bottle of Dermal Cell Basal Medium (485 mL) from cold storage. 
4. Decontaminate the external surfaces of all growth kit component vials and the basal medium bottle by spraying them with 70% ethanol.
5. Using aseptic technique and working in a laminar flow hood or biosafety cabinet, transfer the indicated volume of each growth kit component, as indicated in Table 1, to the bottle of basal medium using a separate sterile pipette for each transfer. 

Table 1. Adult Melanocyte Growth Kit Components 

Component	Volume	Final Concentration
rh Insulin	0.5 mL	5 µg/mL
Ascorbic Acid	0.5 mL	50 µg/mL
L-Glutamine	15.0 mL	6 mM
Epinephrine	0.5 mL	1.0 µM
Calcium Chloride	840 µL	1.5 mM
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 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

6. 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.
7. 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.

1. Obtain one growth kit from the freezer; make sure that the caps of all components are tight.
2. Thaw the components of the growth kit just prior to adding them to the basal medium.
3. Obtain one bottle of Dermal Cell Basal Medium (485 mL) from cold storage.
4. Decontaminate the external surfaces of all growth kit component vials and the basal medium bottle by spraying them with 70% ethanol.
5. Using aseptic technique and working in a laminar flow hood or biosafety cabinet, transfer the indicated volume of each growth kit component, as indicated below, to the bottle of basal medium using a separate sterile pipette for each transfer.

1. Adult Melanocyte Growth Kit Components
   • rh Insulin, 0.5 mL (Final concentration 5 µg/mL)
   • Ascorbic acid, 0.5 mL (Final concentration 50 µg/mL)
   • L-Glutamine, 15.0 mL (Final concentration 6 mM)
   • Epinephrine, 0.5 mL (Final concentration 1.0 µM)
   • Calcium Chloride 840 µL (Final concentration 1.5 mM)
   • Peptide growth factor, 1 mL (0.2% proprietary formulation)
   • M8 supplement, 5 mL (1% proprietary formulation)

Antimicrobials and phenol red are not required for proliferation but may be added if desired. The recommended volume of either of the optional components to be added to the complete growth media is summarized below.


2. Addition of Antimicrobials/Antimycotics and Phenol Red (Optional):
   • Gentamicin-Amphotericin B Solution, 0.5 mL (Final concentration Gentamicin: 10 µg/mL; Amphoteriicin B: 0.25 µg/mL)
   • Penicillin-Streptomycin-Amphotericin B Solution, 0.5 mL (Final concentration Penicillin: 10 Units/mL, Streptomycin: 10 µg/mL, Amphotericin B: 25 ng/mL).
   • Phenol Red, 0.5 mL (Final concentration 33 µM)

6. 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.
7. Complete growth media should be stored in the dark at 2°C to 8°C (do not freeze). When stored under these conditions, complete growth media is stable for 30 days.

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.

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