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modal number = 73; range = 63 to 78. This is a hypertriploid human cell line having the modal chromosome number of 73 occurring in 26% of cells. However, cells having 75 chromosomes also occurred at a high rate （24%）. Higher ploidies were found at 0.4%. At least 16 marker chromosomes are common to all metaphases examined: paired del（2）（p2209） and del（9）（p22） and single der（14）t（1;14）（p22;q22）, del（3）（p2501）, del（7）（q3209）, del（10）（p1301）, t（13q17q）, t（3p13q） and at least eight others. The del（5） （p13）, del（7） （q11） and a few others occurred in some, and many others were seen only once. N7 and N21 occurred in 4 or more copies per cell. Most cells had two X and two Y chromosomes.
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. 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:
Remove and discard culture medium.
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.
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. Subc*tion Ratio: A subc*tion ratio of 1:10 to 1:15 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 30-2002 recommended serum:ATCC 30-2020
References:
22287: Arnstein P, et al. Propagation of human tumors in antithymocyte serum-treated mice. J. Natl. Cancer Inst. 52: 71-84, 1974. PubMed: 4544026 22907: Day RS, Ziolkowski CH. Human brain tumour cell strains with deficient host-cell reactivation of N-methyl-N'-nitro-N-nitrosoguanidine-damaged adenovirus 5. Nature 279: 797-799, 1979. PubMed: 450131
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