| Derivation |
This line was originally thought to be derived from normal conjunctiva, but was subsequently found, based on isoenzyme analysis, HeLa marker chromosomes, and DNA fingerprinting, to have been established via HeLa cell contamination. |
| Comments |
The cells are positive for keratin by immunoperoxidase staining.
This cell line contains HeLa marker chromosomes and has type A G6PD (see Nature 259:211-213, 1976 and In Vitro 14:469-475, 1978).
Note: Cells of this line contain HeLa marker chromosomes, and were derived via HeLa contamination |
| 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.
- Remove and discard culture medium.
- 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.
- 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.
- Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
- Add appropriate aliquots of the cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: 1:4 to 1:10
Medium Renewal: Every 2 to 3 days
Note:For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005. |
| References |
Wong SC, Kilbourne ED. Changing viral susceptibility of a human cell line in continuous cultivation. I. Production of infective virus in a variant of the Chang conjunctival cell following infection with swine or N-WS influenza viruses. J. Exp. Med. 113: 95-110, 1961. PubMed: 13786478
Chang RS. Continuous subcultivation of epithelial-like cells from normal human tissues. Proc. Soc. Exp. Biol. Med. 87: 440-443, 1954. PubMed: 13237268
. . Virology 26: 478, 1965.
Gomez-Duarte OG, et al. Binding of vitronectin to opa-expressing Neisseria gonorrhoeae mediates invastion of HeLa cells. Infect. Immun. 65: 3857-3866, 1997. PubMed: 9284164
St. Geme JW, et al. Characterization of the genetic locus encoding Haemophilus influenzae type b surface fibrils. J. Bacteriol. 178: 6281-6287, 1996. PubMed: 8892830
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.
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