Efficient generation of functional hepatocyte-like cells from mouse liver progenitor cells via indirect co-culture with immortalized human hepatic stellate cells

• Published in: Hepatobiliary & pancreatic diseases international 2016-04, Vol.15 (2), p.173-179
• Main Authors: Pan, Xiao-Ping, Wang, Yi-Ni, Yu, Xiao-Peng, Zhu, Chun-Xia, Li, Jian-Zhou, Du, Wei-Bo, Zhang, Yi-Min, Cao, Hong-Cui, Zhang, Yan-Hong, Zhu, Dan-Hua, Yeoh, George C, Li, Lan-Juan
• Format: Article
• Language: English
• Subjects: Albumins - metabolism; Animals; Biomarkers - metabolism; Cell Differentiation; Cell Line; Cell Shape; Coculture Techniques; Culture Media, Conditioned - metabolism; Cytochrome P-450 CYP1A2 - metabolism; Endocrinology & Metabolism; Gastroenterology and Hepatology; Gene Expression Regulation; Glycogen - metabolism; Hepatic Stellate Cells - metabolism; hepatocytes; Hepatocytes - metabolism; Humans; immortalized human hepatic stellate cells; indirect co-culture; Lipoproteins, LDL - metabolism; Liver - cytology; Liver - metabolism; liver progenitor cells; Male; Mice, Inbred C57BL; Paracrine Communication; Phenotype; Stem Cells - metabolism; Time Factors; Urea - metabolism
• ISSN: 1499-3872
• DOI: 10.1016/S1499-3872(16)60074-7

Background Differentiation of liver progenitor cells (LPCs) to functional hepatocytes holds great potential to develop new strategies for hepatocyte transplantation and the screening of drug-induced cytotoxicity. However, reports on the efficient and convenient hepatic differentiation of LPCs to hepatocytes are few. The present study aims to investigate the possibility of generating functional hepatocytes from LPCs in an indirect co-culture system. Methods Mouse LPCs were co-cultured in Transwell plates with an immortalized human hepatic stellate cell line (HSC-Li) we previously established. The morphology, expression of hepatic markers, and functions of mouse LPC-derived cells were monitored and compared with those of conventionally cultured LPCs. Results Co-culturing with HSC-Li cells induced differentiation of mouse LPCs into functional hepatocyte-like cells. The differentiated cells were morphologically transformed into hepatocyte-like cells 3 days after co-culture initiation. In addition, the differentiated cells expressed liver-specific genes and possessed hepatic functions, including glycogen storage, low-density lipoprotein uptake, albumin secretion, urea synthesis, and cytochrome P450 1A2 enzymatic activity. Conclusions Our method, which employs indirect co-culture with HSC-Li cells, can efficiently induce the differentiation of LPCs into functional hepatocytes. This finding suggests that this co-culture system can be a useful method for the efficient generation of functional hepatocytes from LPCs.