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Cytogenetic Analysis: modal number = 64; range = 60 to 67.
The cell line is aneuploid human, presumably female (X, abnormal X) with most chromosome counts in the hypotriploid range.; Normal chromosomes X, N2, N3, N7, N8, N10, and N22 are clearly under-represented due to their involvement in the formation of the many marker (19) chromosomes present in this cell line.; A normal chromosome N1 (or two) is identified in each karyotype, but, in addition, regions of chromosome N1 are also present in five different marker chromosomes.; Variation is evident in the normal and marker chromosome copy number from karyotype to karyotype.
Isoenzymes: AK-1, 1
ES-D, 1
G6PD, A
GLO-I, 1-2
Me-2, 1-2
PGM1, 1
PGM3, 2
Age: 51 years
Gender: female
Ethnicity: Black
Comments: The MDA-MB-468 cell line was isolated in 1977 by R. Cailleau, et al., from a pleural effusion of a 51-year-old Black female patient with metastatic adenocarcinoma of the breast. Although the tissue donor was heterozygous for the G6PD alleles, the cell line consistently showed only the G6PD A phenotype.There is a G -> A mutation in codon 273 of the p53 gene resulting in an Arg -> His substitution.EGF receptor is present at 1 X 10(6) per cell.
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%.
Atmosphere: air, 100%
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 1:2 to 1:4 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
0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101
Cell culture tested DMSO:ATCC 4-X
References: 1206: Brinkley BR, et al. Variations in cell form and cytoskeleton in human breast carcinoma cells in vitro. Cancer Res. 40: 3118-3129, 1980. PubMed: 7000337
22429: Siciliano MJ, et al. Mutually exclusive genetic signatures of human breast tumor cell lines with a common chromosomal marker. Cancer Res. 39: 919-922, 1979. PubMed: 427779
22544: Pathak S, et al. A human breast adenocarcinoma with chromosome and isoenzyme markers similar to those of the HeLa line. J. Natl. Cancer Inst. 62: 263-271, 1979. PubMed: 283262
22656: Cailleau R, et al. Long-term human breast carcinoma cell lines of metastatic origin: preliminary characterization. In Vitro 14: 911-915, 1978. PubMed: 730202
22930: Nigro JM, et al. Mutations in the p53 gene occur in diverse human tumour types. Nature 342: 705-707, 1989. PubMed: 2531845
22977: Bates SE, et al. Expression of the transforming growth factor-alpha/epidermal growth factor receptor pathway in normal human breast epithelial cells. Endocrinology 126: 596-607, 1990. PubMed: 2294006
23111: Avila MA, et al. Quercetin mediates the down-regulation of mutant p53 in the human breast cancer cell line MDA-MB468. Cancer Res. 54: 2424-2428, 1994. PubMed: 8162591
32275: Littlewood-Evans AJ, et al. The osteoclast-associated protease cathepsin K is expressed in human breast carcinoma. Cancer Res. 57: 5386-5390, 1997. PubMed: 9393764
32467: Zamora-Leon SP, et al. Expression of the fructose transporter GLUT5 in human breast cancer. Proc. Natl. Acad. Sci. USA 93: 1847-1852, 1996. PubMed: 8700847
