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

摘要

BACKGROUND: Differentiation of liver progenitor cells (LPCs) to functional hepatocytes holds great potential to de-velop new strategies for hepatocyte transplantation and the screening of drug-induced cytotoxicity. However, reports on the efifcient 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 differentia-tion 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 addi-tion, the differentiated cells expressed liver-speciifc 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 efifciently induce the differentiation of LPCs into functional hepatocytes. This ifnding suggests that this co-culture system can be a useful method for the efifcient generation of functional hepatocytes from LPCs.