| 產品名稱 |
SYF + c-Src |
| 商品貨號 |
B165808 |
| Organism |
Mus musculus, mouse |
| Tissue |
embryo |
| Cell Type |
fibroblast immortalized with SV40 large T antigen |
| Product Format |
frozen |
| Morphology |
fibroblast |
| Culture Properties |
adherent |
| Biosafety Level |
2 [Cells contain Papovavirus; SV40 viral sequences]
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. |
| Age |
9.5 days gestation |
| Applications |
The SYF + c-Src cell line expresses wild type c-Src and can be
used as a control for SYF cells. |
| Storage Conditions |
liquid nitrogen vapor phase |
| Derivation |
SYF (deficient for Src, Yes, and Fyn) cells were generated in 1997 from mouse embryos harboring functional null mutations in both alleles of the Src family protein tyrosine kinases, Src, Yes, and Fyn. c-Src was reintroduced into a SYF triple mutant cell line via the retroviral vector pLXSH. The cells were immortalized before passage 2 by infection with a retroviral vector transducing the SV40 large T antigen. |
| Comments |
Cells deficient for Src, Yes, and Fyn are available as SYF (ATCC CRL-2459).
The cells are both neomycin and hygromycin resistant. |
| Complete Growth Medium |
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. 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 - 0.53 mM 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 1:6 to 1:12 is recommended
Medium Renewal: Every 2 to 3 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 |
Culture medium, 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC? Catalog No. 4-X. |
| Culture Conditions |
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO 2), 5%
|
| Name of Depositor |
RA Klinghoffer, P Soriano |
| Deposited As |
mouse |
| References |
Klinghoffer RA, et al. Src family kinases are required for integrin but not PDGFR signal transduction. EMBO J. 18: 2459-2471, 1999. PubMed: 10228160
|
