| Karyotype |
model number = 39; range = 38 to 41.
This is a hypodiploid human cell line with the modal chromosome number of 39, occurring in 62% of cells. The rate of cells with higher ploidies was 16%. Sixteen markers were found in single copies in all cells. They include del(2) (q34), del(8) (p21), 14p+, der(X)t(X;14) (q28;q13), t(3q8q) and eleven others. A few different markers, all of which had 11qter--11p13 [der(11)] in common, were found. These markers appeared to be mutually exclusive, each cell having one, and only one, marker at a time. Normal X, N8, N11, N21 and N22 were absent. N1, N10, N15, and N16 were paired, and other normal chromosomes were single. |
| 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: 1:2 to 1:5
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.
|
| References |
Goodfellow M, et al. One hundred and twenty-seven cultured human tumor cell lines producing tumors in nude mice. J. Natl. Cancer Inst. 59: 221-226, 1977. PubMed: 77210034
Pater MM, Pater A. Human papillomavirus types 16 and 18 sequences in carcinoma cell lines of the cervix. Virology 145: 313-318, 1985. PubMed: 2992153
Yee C, et al. Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. Am. J. Pathol. 119: 361-366, 1985. PubMed: 2990217
Hendricks DT, et al. FHIT gene expression in human ovarian, endometrial, and cervical cancer cell lines. Cancer Res. 57: 2112-2115, 1997. PubMed: 9187105
Geisbill J, et al. Detection and characterization of human papillomavirus type 45 DNA in the cervical carcinoma cell line MS751. J. Gen. Virol. 78: 655-658, 1997. PubMed: 9049418
Smith KJ, et al. The APC gene product in normal and tumor cells. Proc. Natl. Acad. Sci. USA 90: 2846-2850, 1993. PubMed: 8385345
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.
|
