| Comments |
This line is a derivative of the original CHO cell line and is the parental line for several glycosylation mutants (see ATCC CRL-1735, 1736 and 1737). The cells are proline auxotrophs and must be grown in a medium that contains proline (40 mg/L). The population contains Pro+ revertants at high frequency (approximately 1 in 250 cells), and should be cloned if it is desired to use the Pro- marker in complementation studies. |
| Subculturing |
Remove medium, add fresh 0.25% trypsin to 0.02% EDTA, let sit for 2 to 3 minutes, remove trypsin solution and let the culture sit at room temperature for 5 to 10 minutes or until the cells detach. Add fresh medium, aspirate and dispense into new flasks. ALTERNATIVELY, cultures also can be maintained by addition or replacement of fresh medium. Start cultures at 2 x 104 cells/mL and maintain between 2 x 104 and 1 x 106 cells/mL.
Subcultivation Ratio: 1:4 to 1:8
Growth Properties: Monolayer or suspension
Medium Renewal: Two to three times weekly |
| References |
Stanley P, et al. Selection and characterization of eight phenotypically distinct lines of lectin-resistant Chinese hamster ovary cells. Cell 6: 121-128, 1975. PubMed: 1182798
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.
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