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Isolation: Isolation date: August, 1971
Virus Susceptibility: Human poliovirus 1
Vesicular stomatitis virus
Reverse Transcript: negative
DNA Profile （STR）: Amelogenin: X
CSF1PO: 11,12
D13S317: 9
D16S539: 12
D5S818: 12
D7S820: 8,10
THO1: 7,8
TPOX: 8
vWA: 17,19
Cytogenetic Analysis: modal number = 63; range = 45 to 65.
Approximay 10% of the cells examined possessed a pair of dicentric chromosomes.
Isoenzymes: G6PD, B
Age: 55 years
Gender: female
Ethnicity: Caucasian
Comments: Electron microscopic studies show many bulb gap junctions （BGJ）.
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Leibovitz's L-15 Medium, Catalog No. 30-2008. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Temperature: 37.0°C
Subculturing: Protocol:
1.Remove and discard culture medium.
2.Briefly rinse the cell layer with 0.25% （w/v） Trypsin- 0.53 mM EDTA solution to remove all traces of serum that 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.
Subc*tion Ratio: A subc*tion ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Preservation: Freeze medium: Complete growth medium supplemented with 5% （v/v） DMSO
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium （without the additional supplements or serum described under ATCC Medium）:ATCC 30-2008
recommended serum:ATCC 30-2020
References: 22140: . . In Vitro 8: 443, 1973.
22526: Leibovitz A, et al. New human cancer cell culture lines. I. SW-13, small-cell carcinoma of the adrenal cortex. J. Natl. Cancer Inst. 51: 691-697, 1973. PubMed: 4765382
22536: Fogh J, et al. Absence of HeLa cell contamination in 169 cell lines derived from human tumors. J. Natl. Cancer Inst. 58: 209-214, 1977. PubMed: 833871
22539: Fogh J, 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: 327080
23212: Lasfargues EY, Ozzello L. C*tion of human breast carcinomas. J. Natl. Cancer Inst. 21: 1131-1147, 1958. PubMed: 13611537
26278: Johnson RG, Sheridan JD. Junctions between cancer cells in culture: ultrastructure and permeability. Science 174: 717-719, 1971. PubMed: 4330805
26279: Pinto da Silva P, Gilula NB. Gap junctions in normal and transformed fibroblasts in culture. Exp. Cell Res. 71: 393-401, 1972. PubMed: 4339896
