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RPMI1640, 52.5%; FBS, 40%; DMSO, 7.5%

RPMI1640 with L-glutamine (300mg/L), 25mM HEPES and 25mM NaHCO3, 90%; heat inactivated fetal bovine serum (FBS), 10%

RPMI1640 with L-glutamine (300mg/L), 25mM HEPES and 25mM NaHCO3, 95%; heat inactivated fetal bovine serum (FBS), 5%

Remove medium, add fresh 0.25 % trypsin 0.02 % EDTA solution, stand culture flask at 37''C for 3 to 5 minutes, add culture medium and collect the cells, transfer the medium into 15ml tube, centrifuge, aspirate the medium, resuspend the pellets with culture medium and dispense into the culture flask

Oncogene : BRCA1 mutation, insertion C at nucleotide 5382, Her2/neu(-), p53(+). This line grows as large epithelial cells with a tendency to float at high cell densities. BRCA1 analysis revealed that the cell line is homozygous for the BRCA1 5382C mutation, whereas the lymphoblastoid cell line derived from the same patient is heterozygous for the same mutation. This mutation was present in tow other family members; an identical sister also developed breast cancer. The cell line has an acquired mutation of TP53 with wild-type allele loss; an acquired homozygous deletion of the PTEN gene, and loss of heterozygosity at multiple loci known to be involved in the pathogenesis of breast cancer. This cell line is positive for the epithelial cell specific marker Epithelial Glycoprotien 2 (EGP2) and for cytokeratin 19. The cells are negative for expression of estrogen receptor (ER) and progesterone receptor (PR).

Remove two thirds of medium and add fresh medium (depending on cell density) every 2 to 3 days