Culturing Human Cell Lines for Transport Studies

The human choriocarcinoma cell lines Jeg-3 and BeWo were purchased from ATCC and grown in Dulbecco’s Modified Eagle’s Medium (DMEM) and Ham’s F12K Kaighn’s Medium, respectively. HEK 293 cell lines transfected with P-gp, ABCG2, or MRP1 were grown in DMEM supplemented with 1 mg/mL geneticin35 (link). Drug-selected H460-MX20 cells that overexpress ABCG2 were grown in RPMI media containing 20 nM mitoxantrone. All media above was supplemented with 10% Fetal Bovine Serum (FBS), penicillin/streptomycin, and 2 mM glutamine. Primary human villous trophoblast (HVT) cells were purchased from Sciencell Research Laboratories and maintained in Trophoblast medium (Sciencell) with included supplements. All cells were incubated at 37 °C in 5% CO_2. Cells were sub-cultured utilizing a 0.25% trypsin, 9 mM EDTA solution, and were used in experiments between passages 4–20.