Primary Dermal Fibroblasts; Normal, Human, Adult (ATCC® PCS-201-012)

Organism: Homo sapiens, human  /  Tissue: skin  / 

Permits and Restrictions

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Organism Homo sapiens, human
Tissue skin
Morphology Spindle-shaped; cells are bipolar and refractile.
Growth Properties Adherent
Biosafety Level 1
Age Adult
Gender Lot-specific
Ethnicity Lot-specific
Applications
Response to pathogens, skin aging, wound healing, gene delivery, skin diseases (e.g., scleroderma); cosmetics research/testing.
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 supports more prolific growth compared to the serum-free medium.
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. 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


  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 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. 
Volume 1 mL
Sterility Tests
Bacteria and Yeasts: Negative.
Mycoplasma: Negative
Viral Testing
Hepatitis B: Negative
Hepatitis C: Negative
HIV: Negative
Viability ≥ 50% when thawed from cryopreservation
Population Doubling Time ≥ 10 doublings in serum-free media
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
FAQs
  1. Transfection of Primary Human Fibroblasts
    Yes, we have tested and recommend transfecting primary uterine fibroblasts with either TransfeX™ Transfection Reagent (ACS-4005) or GeneXPlus™
    Date Updated: 9/15/2014
  2. ATCC primary dermal fibroblasts PDL
    ATCC primary fibroblasts have been tested to reach at least 10 PDLs when maintained according to the product sheet. However, these cells are typically capable of proliferating in culture for long...
    Date Updated: 9/15/2014
  3. Serum-free media for dermal fibroblasts
    Yes, we offer a serum-free growth kit (PCS-201-040) which when combined with our dermal fibroblast basal medium (PCS-201-030) creates a completely defined medium for the serum free culture of hum...
    Date Updated: 9/15/2014
References

Rakowska P, Lamarre B, Ryadnov M. Probing label-free intracellular quantification of free peptide by MALDI-ToF mass spectrometry. Methods 68(2):331-37, 2014. PubMed: 24657280

Malpass G, Arimilli S, Prasad G, Howlett C. Regulation of Gene Expression by Tobacco Product Preparations in Cultured Human Dermal Fibroblasts. Toxicol Appl Pharmacol 279(2): 211-219, 2014. PubMed: 24927667

Malpass G, Arimilli S, Prasad G, Howlett A. Complete artificial saliva alters expression of proinflammatory cytokines in human dermal fibroblasts. Toxicol Sci 134(1):18-25, 2013. PubMed: 23629517

Kim S, Moon S, Lee Y, et al. Alternative xeno-free biomaterials derived from human umbilical cord for the self-renewal ex-vivo expansion of mesenchymal stem cells. Stem Cells Dev 22(22):3025-38, 2013. PubMed: 23786292

Whiteley J, Bielecki R, Li M, et al. An expanded population of CD34+ cells from frozen banked umbilical cord blood demonstrate tissue repair mechanisms of mesenchymal stromal cells and circulating angiogenic cells in an ischemic hind limb model. Stem Cell Rev 10(3):338-50, 2014. PubMed: 24443055

Pandit V, Zuidema J, Venuto K, et al. Evaluation of multifunctional polysaccharide hydrogels with varying stiffness for bone tissue engineering. Tissue Eng Part A 19(21-22):2452-63, 2013. PubMed: 23724786

Golberg A, Bei M, Sheridan R, Yarmush M. Regeneration and control of human fibroblast cell density by intermittently delivered pulsed electric fields. Biotechnol Bioeng 110(6):1759-68, 2013. PubMed: 23297079

Ansari C, Tikhomirov G, Hong S, et al. Development of novel tumor‐targeted theranostic nanoparticles activated by membrane‐type matrix metalloproteinases for combined cancer magnetic resonance imaging and therapy. Small 10(3):566-75, 2014. PubMed: 24038954

Pusztai E, Toulokhonova I, Temple N, et al. Synthesis and photophysical properties of asymmetric substituted silafluorenes. Organometallics 32(9):2529-35, 2013.

Cam M, Bid H, Xiao L, et al. p53/TAp63 and AKT regulate mTORC1 signaling through two independent parallel pathways in the presence of DNA damage. J Biol Chem 289(7):4083-94, 2013. PubMed: 24366874