Hepatocyte culture media kit

Cryopreserved human hepatocytes from three donors (#1: HMC 514, #2: HFC920, #3: HMC1034), the High Viability Cryohepatocyte Recovery Kit, Hepatocyte Culture Media Kit, Matrigel matrix, and collagen I-coated 24-well plates were obtained from Corning Life Sciences (Woburn, MA, USA). GlutaMAX-I supplement was purchased from Gibco (Grand Island, NY, USA). Cryopreserved HepaRG cells, HepaRG Thaw, Plate, and General Purpose Medium Supplement, and HepaRG Induction Medium Supplement were obtained from Biopredic (Rennes, France). Calcitriol, omeprazole, 6-(4-chlorophenyl)imidazo[2,1-b] [1,3]thiazole-5-carbaldehyde O-3,4-dichlorobenzyl) oxime (CITCO), rifampin, bupropion, and William’s Medium E were purchased from Sigma-Aldrich (St. Louis, MO, USA). Superscript III First-Strand Synthesis SuperMix, TRIzol, and diethyl pyrocarbonate (DEPC)-treated water were obtained from Invitrogen (Carlsbad, CA, USA). Quantitative polymerase chain reaction (qPCR) reagents were purchased from Applied Biosystems (Foster City, CA, USA). Dimethyl sulfoxide (DMSO) was obtained from Merck (Merck-Millipore, Darmstadt, Germany). All other chemicals were of reagent grade or better and were used without further purification.

The HepAD38 cell line is derived from HepG2 cells and contains the HBV genome (subtype ayw) under tetracycline control (29 (link)). HepAD38 cells were maintained in Dulbecco modified Eagle medium/F-12 with 10% fetal calf serum (FCS), 3.5 × 10^–7 M hydrocortisone hemisuccinate, and insulin at 5 μg/ml. Primary human hepatocytes (PHHs) were purchased from Corning (catalog number 454541, lot number 399) and maintained in PHH medium (Corning catalog number 355056, Corning hepatocyte culture media kit, 500 ml) as recommended by the manufacturer.