SYF + c-Src (ATCC® CRL-2498)

Organism: Mus musculus, mouse  /  Cell Type: fibroblast immortalized with SV40 large T antigen  /  Tissue: embryo  / 

Permits and Restrictions

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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]
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.
  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 1:6 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 (CO2), 5%
Name of Depositor RA Klinghoffer, P Soriano
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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Restrictions

The line is available with the following restrictions:

  1. This cell line was deposited at the ATCC by Dr. P Soriano and Dr. RA Klinghoffer. The cell line is provided for research purposesonly and is available under the condition that the material will not be used for commercial purposes or distributed it to third parties The cells are provided without warranty of merchantability or fitness for a particular purpose or any other warranty, expressed or implied.
  2. Any proposed commercial use of the these cells, or their products must first be negotiated with the Fred Hutchinson Cancer Research Center,1100 Fairview Avenue North, C2M-027M, Seattle, WA 98109. Telephone (206) 667-6304, FAX (206) 667-4732.

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