Adult Melanocyte Growth Kit (ATCC® PCS-200-042)

Biosafety Level 1
Product Format frozen 1 Kit
Storage Conditions -20°C (or -70°C for long-term storage)
Comments The Adult Melanocyte Growth Kit contains components that when added to Dermal Cell Basal Medium (ATCC PCS-200-030) create a complete low-serum culture environment for adult melanocytes derived from normal human epidermis (e.g., Primary Epidermal Melanocytes, Normal, Human Adult, ATCC PCS-200-013). The complete low serum medium is formulated to support robust adult melanocyte growth in the absence of cholera toxin and phorbol 12-myristate 13-acetate (PMA). No feeder layers, extracellular matrix proteins or other substrares are required.

The final concentration for each component in complete adult melainocyte growth medium is as follows:
  • rh Insulin: 5 µg/mL
  • Ascorbic Acid: 50 µg/mL
  • L-Glutamine: 6 mM
  • Epinephrine: 1.0 µM
  • Calcium chloride: 1.5 mM
  • Peptide Growth Factor: 0.2%, Proprietary formulation
  • M8 Supplement: 1%, Proprietary formulation
Complete Growth Medium
  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. 
  6. 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

    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 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

  7. 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.
  8. 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.
Volume 1 Kit
Sterility Tests
Negative for bacteria, fungi and yeast

Functional Tests Rate of proliferation and morphology
A Certificate of Analysis (COA) is available upon request for each lot of Adult Melanocyte Growth Kit.
C of A
Certificate of Analysis
Certificate of Analysis