CCD-986Sk (ATCC® CRL-1947)

Organism: Homo sapiens, human  /  Cell Type: fibroblast  /  Tissue: skin  / 

Organism Homo sapiens, human
Tissue
skin
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Age 22 years
Gender female
Ethnicity Black
Storage Conditions liquid nitrogen vapor phase
Derivation
The line was established from skin taken from normal breast tissue removed during breast reduction mammoplasty.
Clinical Data
22 years
Black
female
Comments
The cells have been observed to senesce at about 36 population doublings.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Iscove's Modified Dulbecco's Medium, Catalog No. 30-2005. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation
Freeze Medium: Complete growth medium 95%; DMSO, 5%
Storage Temperature: Liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; carbon dioxide (CO2), 5%
STR Profile
Amelogenin: X
CSF1PO: 11,13
D13S317: 9,12
D16S539: 8
D5S818: 11,13
D7S820: 10,11
THO1: 7,9.3
TPOX: 8,11
vWA: 14,17