Hepatocytes :MH vs. ProliHH vs. PHH vs. HLC. Hepatocytes :MH vs. ProliHH vs. PHH vs. HLC

Hepatocytes isolated from DILI patient's liver (#2064) were cultured for a long term using irrMEF and EMUKK-05, and comprehensive gene expression was compared between Puromycin-treated and non-treated groups. In addition, comprehensive gene expression analysis of human mature hepatocytes, primary cultured cells, and ips cell-derived hepatocyte-like cells were performed as controls. Two-condition experiment, Proliferating hepatocytes (ProilHH) vs. puromycin-treated ProliHH. Primary human hepatocytes (PHH) and isolated humen mature hepatocytes (MH) and human iPSC-derived hepatocyte-like cells (HLC) as controls. Overall design: Hepatocytes were isolated from the livers of human DILI patients and frozen after primary culture. The cells were cultured for a long turm of time in MEF feeder and EMUKK-05 medium (ProliHH), and the gene expression changes between the puromycin- and non-puromoted groups were analyzed. Puromycin was used as a selective agent for hepatocytes.

从药物性肝损伤（Drug-Induced Liver Injury, DILI）患者肝脏（样本编号#2064）分离得到的肝细胞，采用irrMEF与EMUKK-05培养基进行长期培养，并比较嘌呤霉素（Puromycin）处理组与未处理组的全基因组表达谱差异。此外，本研究还对人成熟肝细胞、原代培养肝细胞及诱导多能干细胞（induced pluripotent stem cell, iPSC）来源的肝样细胞（hepatocyte-like cells, HLC）开展全基因组表达谱分析作为对照。本实验为双条件对照实验：增殖型原代人肝细胞（ProliHH，Proliferating Hepatocytes）与经嘌呤霉素处理的ProliHH；并以原代人肝细胞（Primary Human Hepatocytes, PHH）、分离得到的人成熟肝细胞（Mature Hepatocytes, MH）及人诱导多能干细胞来源的肝样细胞（HLC）作为对照。总体实验方案：从药物性肝损伤患者肝脏中分离肝细胞，经原代培养后冻存。后续将细胞置于小鼠胚胎成纤维细胞（Mouse Embryo Fibroblasts, MEF）饲养层及EMUKK-05培养基中进行长期培养（即ProliHH细胞），并分析嘌呤霉素处理组与未处理组的基因表达变化。嘌呤霉素作为肝细胞的筛选试剂使用。