Embryoid-body cells derived from a mouse embryonic stem cell line show differentiation into functional hepatocytes

Abstract

Embryonic stem (ES) cells have a potential to differentiate into various progenitor cells.Here we investigated the differentiation capacity of mouse ES cells into hepatocytes both in vitro and in vivo. During the culture of embryoid bodies (EBs) derived from ES cells, albumin (ALB) messenger RNA (mRNA) was expressed within 12 days after removal of leukemia inhibitory factor, and ␣-fetoprotein (AFP) mRNA was observed within 9 days without additional exogenous growth factors. In ES cells and early EBs, by contrast, neither ALB mRNA nor AFP mRNA was observed. ALB protein was first detected at day 15 and the level increased with the culture period. The differentiation of EBs facilitated the synthesis of urea with the culture period, whereas early EBs and ES cells produced no urea. These results suggest that cultured EBs contain hepatocytes capable of producing ALB and urea. ES cells and the isolated cells from EBs were transplanted through portal vein to the liver after 30% partial hepatectomy of female mice pretreated with 2-acetylaminofluorene. Four weeks after transplantation with isolated cells from day-9 EBs, ES-derived cells containing Y-chromosome in the liver were positive for ALB (0.2% of total liver cells), whereas teratoma was found in mice transplanted with ES cells or EBs up to day 6. The incidence of teratoma was decreased with the culture duration and no teratoma was observed in the liver transplanted with isolated cells from day-9 EBs. In conclusion, our in vitro and in vivo experiments revealed that cultured EBs contain functional hepatocytes or hepatocyte-like cells. (HEPATOLOGY 2002;36:22-29.)

Embryonic development of the liver†

“…The information that has been generated by the study of hepatic development is also now being used in the production of hepatocytes and other liver cells from progenitor cell populations and embryonic stem cells. 124,[153][154][155] For example, Teratani and colleagues demonstrated that embryonic stem cells could be induced to generate cells with hepatic gene expression profiles and functions using a differentiation procedure that included culture with FGF, hepatocyte growth factor, and oncostatin M. 156 The ability to efficiently produce hepatocytes from progenitor or embryonic stem cells is exciting, because such cells could potentially be used for hepatocyte transplantation, gene therapy, the generation of artificial liver devices, or even tissue engineering. [157][158][159] By building on our understanding of the basic mechanisms that lead Fig.…”

Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines

“…that ES cells can be differentiated into hepatocyte-like cells [1][2][3][4][5][6] . ES cell-derived hepatocytes would be useful as a source of material for cell transplantation, bioartificial liver (BAL) support devices, studies on hepatocyte biology and hepatitis viruses and for drug testing.…”

Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver

“…Several studies have reported the induction of ES cell differentiation into hepatocytes both in vitro [6][7][8][9] and in vivo. 10 The in vitro approaches involve the formation of embryoid bodies (EBs) to re-create the inductive microenvironment required for liver organogenesis 11,12 or treatment with specific growth factors and cytokines critical for hepatocyte differentiation such as hepatocyte growth factor, fibroblast growth factor, and oncostatin. 1,13 Also, the introduction of genes promoting endodermal differentiation into ES cells, 14 modification of culture microenvironment by supplementation with extracellular matrix proteins or coculture with other cell types, 15,16 is used to direct ES cells toward a hepatocytic lineage.…”