| Karyotype |
This is a hyperdiploid human cell line. The modal chromosome number is 52, occurring at 40% with polyploidy at 3.8%., Over 2/3 of the chromosome complement consist of those with structural rearrangements (or markers) including 1q, t(1p,16q), del(3)(p13), t(5;14)(q35;q11), t(9q;?)1, t(9q;?)2, 12p+, 20p+, t(Xq,7p)., Twenty-one other markers are common to most cells. The normal chromosomes 1,3,5,9,14,15 and X are absent. All three E group chromosomes are paired., Y chromosome not detected by Q staining. |
| 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 1:6
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.
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| References |
Lieblich JM, et al. Ectopic and eutopic secretion of chorionic gonadotropin and its subunits in vitro: Comparison of clonal strains from carcinomas of lung and placenta. J. Natl. Cancer Inst. 56: 911-917, 1976. PubMed: 62839
Rabson AS, et al. Production of human chorionic gonadotropin in vitro by a cell line derived from a carcinoma of the lung. J. Natl. Cancer Inst. 50: 669-674, 1973. PubMed: 4708151
Dahiya R, et al. Mucin synthesis and secretion in various human epithelial cancer cell lines that express the MUC-1 mucin gene. Cancer Res. 53: 1437-1443, 1993. PubMed: 8443822
Tashjian AH Jr., et al. Subunits of human chorionic gonadotropin: unbalanced synthesis and secretion by clonal cell strains derived from a bronchogenic carcinoma. Proc. Natl. Acad. Sci. USA 70: 1419-1422, 1973. PubMed: 4514312
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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