SV40LT-SMC Clone HEP-SA (ATCC® CRL-2018)

Organism: Rattus norvegicus, rat  /  Cell Type: SV40 transfected  /  Tissue: aorta; smooth muscle  / 

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Organism Rattus norvegicus, rat
Tissue aorta; smooth muscle
Cell Type SV40 transfected
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2  [Cells contain polyomavirus DNA 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 3 months
Strain Sprague-Dawley
Applications
This is a clone of the SV40LT-SMC cell line derived from primary abdominal aortic tissue from young rats.
Transfection was done using conditioned medium from psiTEX cells (which produce a helper free replication defective retrovirus that carries the sequences for large T antigen and neomycin (G418) resistance).
Storage Conditions liquid nitrogen vapor phase
Derivation
This is a clone of the SV40LT-SMC cell line derived from primary abdominal aortic tissue from young rats.
The parental line was created by immortalizing the primary cells by transfection with SV40 large T antigen.
Genes Expressed
alpha actin; large T antigen
Cellular Products
alpha actin; large T antigen
Comments
This is a clone of the SV40LT-SMC cell line derived from primary abdominal aortic tissue from young rats.
The parental line was created by immortalizing the primary cells by transfection with SV40 large T antigen.
Transfection was done using conditioned medium from psiTEX cells (which produce a helper free replication defective retrovirus that carries the sequences for large T antigen and neomycin (G418) resistance).
Clone HEP-SA cells maintain smooth muscle cell characteristics including smooth cell alpha actin expression and growth inhibition by heparin.
Large T antigen is detectable in the nuclei.
Complete Growth Medium The base medium for this cell line is ATCC-formulated DMEM Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: 10% Bovine Calf Serum and 200 mcg/mL G418
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: 1:3 to 1:6
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 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

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 CF Reilly, R McFall
References

Reilly CF. Rat vascular smooth muscle cells immortalized with SV40 large T antigen possess defined smooth muscle cell characteristics including growth inhibition by heparin. J. Cell. Physiol. 142: 342-351, 1990. PubMed: 2154504

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm

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

Reilly CF. Rat vascular smooth muscle cells immortalized with SV40 large T antigen possess defined smooth muscle cell characteristics including growth inhibition by heparin. J. Cell. Physiol. 142: 342-351, 1990. PubMed: 2154504

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.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm