Cell Line:                              BT-474

Cell Description:                 Human mammary gland

 

Culture medium:                      RPMI 1640 medium with 4,5 g/l glucose, sodium bicarbonate, 2 mM L-glutamine, 1,0 mM sodium pyruvate, 10 mM HEPES, 1,2 mM oxaloacetic acid, 0,02 mg/ml bovine insulin, 10 % NCTC 135 medium and 10 % fetal bovine serum.

Alternatively, DMEM:Ham«s F12 (1:1 mixture) supplemented with 2 mM L-glutamine, 1,0 mM sodium pyruvate and 10 % fetal bovine serum may be used.

 

Subculturing:                          Remove medium, rinse with fresh 0,025 % trypsin / 0,02% EDTA solution, remove trypsin and let the culture sit at room temperature (or at 37 ”C) until the cells detach. Add fresh medium, remove trypsin by centrifugation, and dispense into new flasks. Subculture every 6 to 8 days.

Split ratio:                                A ratio of 1:2 to 1:3 is recommended

Fluid Renewal:                        2 to 3 times weekly

Morphology:                            epithelial

Growth Properties:                 Monolayer; the cells form compact, multilayered colonies; growing slowly and rarely becoming confluent.

Description:                            The BT-474 line was isolated by E. Lasfargues and W. G. Coutinho from a solid, invasive ductal carcinoma of the breast.

Tumorigenic:                           yes, in nude mide

Karyotype:                              mode = 55; range = 50 to 112; bimodal shift 58 Š 59 and 100 in later passages with 3 marker chromosomes

Isoenzymes:                           Me-2, 0; PGM3, 1; PGM1, 1; ES-D, 1; AK-1, 1; G6PD, B; GLO-1, 1; Phenotype Frequency Product: 0.0426

Virus Susceptibility:                mouse mammary tumor virus (RIII-MuMTV)

Comments:                              Derived from a 60 years old caucasian female.

Sterility:                                   Tests for mycoplasma, bacteria and fungi were negative

Biosafety Level:                     1

References:                          

Lasfargues EY et al. Isolation of two human tumor epithelial cell lines from solid breast carcinomas. J. Natl. Cancer Inst. 61: 967-978, 1978. Lasfargues EY et al. A human breast tumor cell line (BT-474) that supports mouse mammary tumor virus replication. In Vitro 15: 723-729, 1979. Kumar R, Shepard HM and Mendelsohn J. Regulation of phosphorylation of the c-erbB-2/HER2 gene product by a monoclonal antibody and serum growth factor(s) in human mammary carcinoma cells. Mol Cell Biol 11 (2): 979-986, 1991.