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modal number = 76; range = 45 to 89.
Karyotype unstable within hyperdiploid stemline range 73-78. Ninety percent of the cells examined have 2 or 3 large chromosomes with subterminal centromeres and 1-6 microchromosomes. Secondary constrictions were observed in 59% of the cells, 21% had breaks.
Isoenzymes:
G6PD, B
Age:
3 months gestation
Gender:
female
Ethnicity:
Caucasian
Propagation:
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
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.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:10 is recommended Medium Renewal: Twice 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 30-2003
recommended serum:ATCC 30-2020
parental cell line:ATCC CCL-75
References:
23133: Girardi AJ, et al. SV40 transformation of human diploid cells. A parallel study of viral and karyologic parameters. Ann. Med. Exp. Biol. Fenn. 44: 242-254, 1966. PubMed: 4290661 26230: Jensen F, et al. Autologous and homologous implantation of human cells transformed in vitro by simian virus 40. J. Natl. Cancer Inst. 32: 917-937, 1964. 32701: Wang S, Vos JM. A hybrid herpesvirus infectious vector based on epstein-barr virus and herpes simplex virus type 1 for gene transfer into human cells in vitro and in vivo. J. Virol. 70: 8422-8430, 1996. PubMed: 8970963
