Colo205 ccl 222

Four cell lines were used in this study: the doxorubicin-sensitive Colo205 (CCL-222, ATCC, Manassas, VA, USA) human colon adenocarcinoma cell line; the doxorubicin-resistant Colo320/MDR-LRP expressing P-glycoprotein (P-gp, ABCB1) (MDR1)-LRP (CCL-220.1, ATCC) human colon adenocarcinoma cell line; the hormone-responsive MCF-7 breast cancer cell line and the normal MRC-5 human embryonal lung fibroblast cell line (CCL-171, ATCC). The human cancer cell lines were purchased from LGC Promochem (Teddington, UK), and the MRC-5 cell line is a Sigma-Aldrich (Merck KGaA, Darmstadt, Germany) product. The Colo205 and Colo320 cell lines were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS), 2 mM l-glutamine, 1 mM Na-pyruvate and 10 mM HEPES. The MCF-7 and MRC-5 cells were cultured in Eagle's Minimal Essential Medium (EMEM) containing 4.5 g L⁻¹ of glucose and supplemented with a non-essential amino acid mixture, a selection of vitamins and 10% FBS. All cell lines were incubated at 37 °C, 5% CO₂, 95% air atmosphere. The cells were detached with Trypsin-Versene (EDTA) solution for 5 min at 37 °C.

Media components were obtained from Sigma-Aldrich unless stated otherwise. COLO205 (CCL-222^®) and MDA-MB-231 (HTB-26^®) were purchased from ATCC (Gaithersburg, MD, USA) and authenticated using short-tandem repeat genetic profiling yielding a 100% match to the ATCC database. Cells were used within ten passages from the original stocks and were tested regularly for mycoplasma. COLO205 and MDA-MB-231 cells were cultured in RPMI-1640 and DMEM medium, respectively, supplemented with 10% fetal bovine serum (Life Technologies, Carlsbad, CA, USA) and 2 mmol/L L-glutamine at 37 °C and 5% CO2.