3A-sub E [post crisis of 3A(tPA-30-1)] (ATCC® CRL-1584)

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
Culture Properties adherent
Biosafety Level 2 Cells contain polyomavirus DNA sequences
Applications
This line was established from ATCC CRL-1583 cells that had begun to senesce.
Storage Conditions liquid nitrogen vapor phase
Derivation
This line was established from ATCC CRL-1583 cells that had begun to senesce.
Genes Expressed
at 40C the cells produce human chorionic gonadotropin (hCG)
Comments
This line was established from ATCC CRL-1583 cells that had begun to senesce.
By passage 20 the cells had recovered and appeared capable of unlimited proliferation.
The saturation density (6 X 10 exp5 cells/sq cm) is increased and the morphology is altered relative to the parental line.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. 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-0.53mM 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 33°C.

Subcultivation Ratio: 1:2
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: 33°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 MI Cour
Passage History
By passage 20 the cells had recovered and appeared capable of unlimited proliferation.
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.

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.