Raszip 6 (ATCC® CRL-1917)

Organism: Mus musculus, mouse  /  Tissue: embryo  / 

Permits and Restrictions

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Organism Mus musculus, mouse
Tissue embryo
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.

Age embryo
Strain NIH/Swiss
Applications
This line was established by G.P. Dotto, et al.
The cells produce a high titer of helper free retrovirus carrying the vHa-ras oncogene and the G418 resistance gene.
Storage Conditions liquid nitrogen vapor phase
Derivation
This line was established by G.P. Dotto, et al. in 1985 by transfecting psi2 cells with the raszip 6 DNA construct.
Comments
This line was established by G.P. Dotto, et al. in 1985 by transfecting psi2 cells with the raszip 6 DNA construct.
The cells produce a high titer of helper free retrovirus carrying the vHa-ras oncogene and the G418 resistance gene.
Complete Growth Medium Dulbecco's modified Eagle's Medium with 4.5 g/L glucose and 0.5 mg/ml G418, 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.
  1. Remove and discard culture medium.
  2. 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.
  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: 1:2 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%
Name of Depositor RA Weinberg
Deposited As Mus musculus
Year of Origin 1985
References

Dotto GP, et al. Specific growth response of ras-transformed embryo fibroblasts to tumour promoters. Nature 318: 472-475, 1985. PubMed: 4069218

Mann R, et al. Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell 33: 153-159, 1983. PubMed: 6678608

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

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
References

Dotto GP, et al. Specific growth response of ras-transformed embryo fibroblasts to tumour promoters. Nature 318: 472-475, 1985. PubMed: 4069218

Mann R, et al. Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell 33: 153-159, 1983. PubMed: 6678608

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.