Zell shield

HeLa, MDA-MB-231, HT-29 and MCF-10A cell lines originate from ATCC (Manassas, VA, USA), and were maintained at 37°C in a humidified atmosphere with 5% CO₂ in DMEM medium (Lonza, Basel, Switzerland) supplemented with 10% fetal bovine serum (Dutscher, Dernolsheim, France), 1% Zell Shield (Dutscher, Bernolsheim, France) and 1% non-essentials amino-acids (Lonza, Basel, Switzerland). MCF-10A were maintained in MEBM medium (Lonza, Basel, Switzerland) supplemented with MEGM (Lonza, Basel, Switzerland). All cell lines culture media were added with 1% Zell Shield (Dutscher, Bernolsheim, France).

Cell lines MDA-MB-231 from triple-negative breast cancer and HeLa from cervical cancer were obtained from the American Type Culture Collection (ATCC; Rockville, MD, USA). They were cultured at 37 °C with 5% CO₂ in DMEM middle (Lonza, Basel, Switzerland), supplemented with 10% fetal bovine serum (Dutscher, Brumath, France), 1% Zell Shield (Dutscher) and 1% non-essentials amino-acids (Lonza). Cancer cells were plated in 96-well plates (2 × 10³ cells/wells) in triplicate, incubated at 37 °C with 5% CO₂ for 24 h, and treated with 1,4-Dihydroquinolines for 72 h at different concentrations. Cell viability was determined using CellTiter 96^® AQueous One Solution Cell Proliferation Assay (MTS, Promega, Madison, WI, USA). 20 µL reagent was added in each well and cells were incubated for 2 h at 37 °C/5% CO₂. Absorbance was measured at 490 nm (SPECTROstar Nano, BMG LABTECH, Ortenberg, Germany). Calculations of IC50 were performed using GraphPad Prism V7.0 software (GraphPad Software, La Jolla, CA, USA).

Human cell lines from cervix cancer (HeLa), breast cancer (MCF-7), triple-negative breast cancer (MDA-MB-231), colorectal cancer (HT-29), prostate cancer (DU-145), and a human breast epithelial cell line, arguably the most commonly used normal breast cell model (MCF-10A), were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were cultured at 37 °C in a humid atmosphere containing 5% CO₂, in a DMEM culture medium supplemented with 10% fetal bovine serum (Dutscher, Dernolsheim, France), 1% Zell Shield (Dutscher), and 1% non-essential amino acids (Lonza, Basel, Switzerland). MCF-10A cells were maintained in MEBM (Lonza) supplemented with MEGM (Lonza) and 1% Zell Shield.