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This is a hypertriploid human cell line. The modal chromosome number was 80, occurring in 42% of cells. Cells with 78 chromosomes also occurred at a high frequency. The rate of cells with higher ploidies was 2.5%. There were 30 or more marker chromosomes in each cell. They included der(1)t(1;?) (p32;?), ?t(1p5p), i(3q), t(7q14q), ?t(2p3p) and others. The X and N3 had single copy per cell, and others were generally two to three copies per cell.
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. 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 Atmosphere: air, 95%; carbon dioxide (CO2), 5%
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 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: A subcultivation ratio of 1:4 to 1:10 is recommended Medium Renewal: 2 to 3 times per week
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2003
recommended serum:ATCC 30-2020
References:
25065: Bellet D, et al. Malignant transformation of nontrophoblastic cells is associated with the expression of chorionic gonadotropin beta genes normally transcribed in trophoblastic cells. Cancer Res. 57: 516-523, 1997. PubMed: 9012484
26153: Grossman HB, et al. Improved growth of human urothelial carcinoma cell cultures. J. Urol. 136: 953-959, 1986. PubMed: 3761468
