Dermal Fibroblast Normal; Human, Neonatal (ATCC® PCS-201-010)

Organism: Homo sapiens, human  /  Tissue: Foreskin  /  Cell Type: Fibroblasts

Permits and Restrictions

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Organism Homo sapiens, human
Tissue Foreskin
Cell Type Fibroblasts
Morphology Spindle-shaped; cells are bipolar and refractile.
Growth Properties Adherent
Biosafety Level 1
Disease Normal
Age Neonatal
Gender Male
Ethnicity Batch-specific
Applications
Response to pathogens, skin aging, wound healing, gene delivery, skin diseases (e.g., scleroderma)
Product Format frozen 1 mL
Storage Conditions -130°C or below
Comments
Serum-free medium supports excellent growth curves and normal morphology, as well as serum-free (not animal-free) experimental conditions. The presence of 2% fetal bovine serum in the Fibroblast Growth Kit-Low serum (ATCC PCS-201-041) supports more prolific growth.
Complete Growth Medium
  1. Obtain one growth kit from the freezer; make sure that the caps of all containers are tight.
  2. 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.
  3. Obtain one bottle of Fibroblast Basal Medium (480 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 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.
  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.

 

Table 1. If using the Fibroblast Growth Kit–Serum-Free (ATCC® PCS-201-040), add the indicated volume for each component in the order shown. 

    Component

     

     

    Volume

     

     

    Final Concentration

     

     

    L-glutamine

     

     

    18.75 mL

     

     

    7.5 mM

     

     

    Hydrocortisone Hemisuccinate

     

     

    0.5 mL

     

     

    1 µg/mL

     

     

    HLL Supplement

     

     

    1.25 mL

     

     

    HSA 500 µg/mL

    Linoleic Acid 0.6 µM

    Lecithin 0.6 µg/mL

     

     

    rh FGF b

     

     

    0.5 mL

     

     

    5 ng/mL

     

     

    rh EGF / TGF  b-1 Supplement

     

     

    0.5 mL

     

     

    5 ng/mL

    30 pg/mL

     

     

    rh Insulin

     

     

    0.5 mL

     

     

    5 µg/mL

     

     

    Ascorbic acid

     

     

    0.5 mL

     

     

    50 µg/mL

     

     

     

           Table 2. If using the Fibroblast Growth Kit–Low Serum (ATCC® PCS-201-041), add the indicated volume for each of the following components:

    Component

     

     

    Volume

     

     

    Final Concentration

     

     

    rh FGF b

     

     

    0.5 mL

     

     

    5 ng/mL

     

     

    L-glutamine

     

     

    18.75 mL

     

     

    7.5 mM

     

     

    Ascorbic acid

     

     

    0.5 mL

     

     

    50 µg/mL

     

     

    Hydrocortisone Hemisuccinate

     

     

    0.5 mL

     

     

    1 µg/mL

     

     

    rh Insulin

     

     

    0.5 mL

     

     

    5 µg/mL

     

     

    Fetal Bovine Serum

     

     

    10.0 mL

     

     

    2%

     

     

     

    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 3.

     

    Table 3. 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

     

     

     

    Volume 1 mL
    Sterility Tests
    Bacteria: Negative
    Yeast: Negative
    Mycoplasma: Negative
    Viral Testing
    Hepatitis B: Negative
    Hepatitis C: Negative
    HIV: Negative
    Viability ≥ 50% when thawed from cryopreservation
    Population Doubling Capacity ≥ 10 in complete growth medium
    C of A
    Certificate of Analysis
    Certificate of Analysis
    Notice: Necessary PermitsPermits

    These permits may be required for shipping this product:

    • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
    Basic Documentation