EMT6/AR1

Drug-resistance studies

EMT6/AR1.0 was derived from the mouse mammary tumour cell line EMT6/P (Sigma Catalogue number. 96042344) by growth in increasing amounts of doxorubicin over a period of 8 weeks. The cell line showed a 40-fold greater resistance to doxorubicin, which is also known as adriamycin. The line was reported to express a classical multi-drug resistance (MDR) phenotype with hyperexpression of membrane P-glycoprotein and a spectrum of cross-resistance e.g. against vincristine and colchicine. Overexpression of CP22 (sorcin) and mdr 1a gene was reported. Cells were shown to have a reduced intracellular drug accumulation and exhibit a doubling time of 12-15 hours. It is recommended to culture the cells without drug after resuscitation until the first passage.

Mouse mammary, drug-resistant

MEM (EBSS) + 2mM Glutamine + 1% Non Essential Amino Acids (NEAA) +1.0 μg/ml doxorubicin + 10% Foetal Bovine Serum (FBS).

Split sub-confluent cultures (70-80%) 1:3 to 1:6 using 0.25% trypsin or trypsin/EDTA; 5% CO₂; 37ºC. Resuscitate in drug-free medium, add doxorubicin after first passage.

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