Culturing Liver and Bone Marrow Cells

Hep G2 cells, HuH-7cells, human liver vascular endothelial (TMNK-1) cells and human bile duct epithelial (MMNK-1) cells were obtained from the Japanese Center Research Bank and cultured in Dulbecco’s modified Eagle’s medium (DMEM; Wako) supplemented with 10% foetal bovine serum (Corning) and 100 units/ml of penicillin-streptomycin (Wako). Cells were grown in an incubator at 37 °C and supplied with 5% CO₂. Mouse bone marrow cells were isolated from the femurs and tibias of 8-week-old C57BL/6NcrSlc male mice (Japan SLC) using previously described method^{24 (link)}. Isolated cells were cultured in DMEM (containing 10% fetal bovine serum and 100 units/ml of penicillin-streptomycin) at 37 °C in a 5% CO₂ incubator. All animal experiments conformed to the Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Committees of Laboratory Animal Experimentation (Animal Research Center of Yokohama City University, Yokohama, Japan).