| 產(chǎn)品名稱 |
CCD-1074Sk |
| 商品貨號 |
B164156 |
| 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 |
42 years |
| Gender |
female |
| Applications |
The line was established from skin taken from normal breast tissue removed at mastectomy. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
The line was established from skin taken from normal breast tissue removed at mastectomy. |
| Clinical Data |
female |
| Comments |
The line was established from skin taken from normal breast tissue removed at mastectomy. Cells senesce after approximately 26 population doublings. |
| Complete Growth Medium |
Iscove's modified Dulbecco's medium, 90%; fetal bovine serum, 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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25%Trypsin-0.53mM EDTA solution.
- 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: 1:3 to 1:6
Medium Renewal: Every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005. |
| Cryopreservation |
Complete growth medium described above supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| Culture Conditions |
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5% |
