| 產品名稱 |
SYF |
| 商品貨號 |
B165807 |
| 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 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 days gestation |
| Applications |
This cell line is a suitable transfection host. |
| 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.
The cells were immortalized before passage 2 by infection with a retroviral vector transducing the SV40 large T antigen. SYF cells also carry a gene for neomycin resistance.
|
| Oncogene |
src -; yes -; fyn - |
| Comments |
Src family kinases have been implicated as key regulators of ligand-induced cellular responses including proliferation, survival, adhesion and migration.
This is the first cell line isolated that lacks all three ubiquitously expressed Src family members.
SYF cells display reduced mobility in wound healing and haptotaxis assays but not in platelet derived growth factor (PDGF) induced chemotaxis. |
| 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 that 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
Subculture Ratio: 1:10 to 1:20
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% |
| Culture Conditions |
Temperature: 37°C |
| Name of Depositor |
RA Klinghoffer, P Soriano |
| Deposited As |
mouse |
| Passage History |
The cells were immortalized before passage 2 by infection with a retroviral vector transducing the SV40 large T antigen. SYF cells also carry a gene for neomycin resistance. |
| 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
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