產(chǎn)品名稱 |
CCD-1059Sk |
商品貨號(hào) |
B164150 |
Organism |
Homo sapiens, human |
Tissue |
skin |
Cell Type |
fibroblast |
Product Format |
frozen |
Morphology |
fibroblast |
Culture Properties |
adherent |
Biosafety Level |
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
Disease |
normal |
Age |
20 years |
Gender |
female |
Ethnicity |
Caucasian |
Storage Conditions |
liquid nitrogen vapor phase |
Derivation |
The line was established from skin taken from normal breast tissue removed at biopsy. |
Clinical Data |
20 years
female
Caucasian |
Comments |
Cells senesce after approximately 35 population doublings. |
Complete Growth Medium |
The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. 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 used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
- Remove and discard culture medium.
- Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: Every 3 to 4 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.
|
Cryopreservation |
Complete growth medium, 95%; DMSO, 5% |
Culture Conditions |
Temperature: 37°C |
STR Profile |
Amelogenin: X CSF1PO: 11,12 D13S317: 12 D16S539: 9,11 D5S818: 11 D7S820: 11,12 THO1: 9,9.3 TPOX: 8 vWA: 16,18 |
Isoenzymes |
G6PD, B |
Population Doubling Capacity |
Cells senesce after approximately 35 population doublings. |