CHO 1-15 [subscript 500] (ATCC® CRL-9606)

Organism: Cricetulus griseus, hamster, Chinese  /  Tissue: ovary  /  Disease: normal

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Organism Cricetulus griseus, hamster, Chinese
Tissue ovary
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease normal
Age adult
Gender female
Shipping Information Distributed: Frozen
Storage Conditions liquid nitrogen vapor phase
Derivation
The line was produced by transfecting CHO cells with a plasmid (pETPFR, ATCC 40403) to produce a system in which human tissue plasminogen activator is produced. 
 
HeLa Markers N
Genes Expressed
human tissue plasminogen activator (tPA, t-PA)
Comments

Expresses human tissue plasminogen activator (t-PA) in eukaryotic DHFR deficient cell lines grown in selective media (glycine, hypoxanthine, thymidine deficient). (Goeddel DV, et al. Human tissue plasminogen activator. US Patent 4,766,075 dated Aug 23 1988)

Contains a EcoRI/BamHI fragment of about 2.2 kb containing t-PA and a 1.7 kb SacII fragment containing wild-type dihydrofolate reductase under SV40 control. (Goeddel DV, et al. Human tissue plasminogen activator. US Patent 4,766,075 dated Aug 23 1988)

Complete Growth Medium Ham's F12 medium, 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: 1:5 to 1:10
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 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 Genentech, Inc.
References

Goeddel DV, et al. Human tissue plasminogen activator. US Patent 4,766,075 dated Aug 23 1988

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

Goeddel DV, et al. Human tissue plasminogen activator. US Patent 4,766,075 dated Aug 23 1988

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