3A(tPA-30-1) (ATCC® CRL-1583)

Organism: Homo sapiens, human  /  Cell Type: SV40 transformed  /  Tissue: placenta  / 

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Organism Homo sapiens, human
Tissue placenta
Cell Type SV40 transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2  [Cells contain polyomavirus DNA sequences]
Applications
The cells express the transformed phenotype at the permissive temperature (33C) and the non-transformed phenotype at the non-permissive temperature (40C).
Storage Conditions liquid nitrogen vapor phase
Karyotype This is a hypodiploid human cell line with the modal chromosome number of 42 occurring in 34% of cells. However, cells with 44 chromosomes also occurred at a high frequency (26%). Most cells contained 41-45 chromosomes. Cells having a higher ploidy occurred at 20.7%, which is very high. Most cells averaged five or six marker chromosomes, none of which appeared twice in the cells examined. No chromosomes were paired consistently.
Genes Expressed
At 40°C the cells synthesize human chorionic gonadotropin (hCG) and alkaline phosphatase
Comments
The cells are transformed by SV40 ts30.
The cells express the transformed phenotype at the permissive temperature (33C) and the non-transformed phenotype at the non-permissive temperature (40C).
The line has a limited life expectancy of 15 to 18 passages before entering the crisis stage.
Complete Growth Medium Alpha minimum essential medium without ribonucleosides and deoxyribonucleosides, 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- 0.53 mM EDTA solution to remove all traces of serum that 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 33°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 33°C to 34°C. Normal placental functions are expressed when the cells are incubated at the restrictive temperature (40°C).

Subcultivation Ratio: 1:2 to 1:4
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%
STR Profile
Amelogenin: X
CSF1PO: 10,12
D13S317: 11
D16S539: 9,12
D5S818: 11,13
D7S820: 10
THO1: 9,9.3
TPOX: 8,11
vWA: 16,17
Name of Depositor J Chou
Passage History
The line has a limited life expectancy of 15 to 18 passages before entering the crisis stage.
References

Chou JY. Human placental cells transformed by tsA mutants of simian virus 40: a model system for the study of placental functions. Proc. Natl. Acad. Sci. USA 75: 1409-1413, 1978. PubMed: 206898

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

Chou JY. Human placental cells transformed by tsA mutants of simian virus 40: a model system for the study of placental functions. Proc. Natl. Acad. Sci. USA 75: 1409-1413, 1978. PubMed: 206898

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