Derivation and Characterization of Hepatic Progenitor Cells from Human Embryonic Stem Cells

Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, College of Life Sciences, Peking University, Beijing, China.
PLoS ONE (Impact Factor: 3.23). 02/2009; 4(7):e6468. DOI: 10.1371/journal.pone.0006468
Source: PubMed


The derivation of hepatic progenitor cells from human embryonic stem (hES) cells is of value both in the study of early human liver organogenesis and in the creation of an unlimited source of donor cells for hepatocyte transplantation therapy. Here, we report for the first time the generation of hepatic progenitor cells derived from hES cells. Hepatic endoderm cells were generated by activating FGF and BMP pathways and were then purified by fluorescence activated cell sorting using a newly identified surface marker, N-cadherin. After co-culture with STO feeder cells, these purified hepatic endoderm cells yielded hepatic progenitor colonies, which possessed the proliferation potential to be cultured for an extended period of more than 100 days. With extensive expansion, they co-expressed the hepatic marker AFP and the biliary lineage marker KRT7 and maintained bipotential differentiation capacity. They were able to differentiate into hepatocyte-like cells, which expressed ALB and AAT, and into cholangiocyte-like cells, which formed duct-like cyst structures, expressed KRT19 and KRT7, and acquired epithelial polarity. In conclusion, this is the first report of the generation of proliferative and bipotential hepatic progenitor cells from hES cells. These hES cell-derived hepatic progenitor cells could be effectively used as an in vitro model for studying the mechanisms of hepatic stem/progenitor cell origin, self-renewal and differentiation.

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Available from: Dongxin Zhao, Mar 13, 2014
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    • "It has been reported that LPC-like cells were established from hESCs/hiPSCs (Takayama et al., 2013; Yanagida et al., 2013; Zhao et al., 2009), and these cells were shown to proliferate and differentiate into hepatocyte-like cells or cholangiocyte-like cells. These LPCs were either isolated by cell sorting using a combination of specific cell surface markers or generated by adenovirus-mediated gene transfer to promote hepatic lineage differentiation. "
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    ABSTRACT: To develop a culture system for large-scale production of mature hepatocytes, liver progenitor cells (LPCs) with a high proliferation potential would be advantageous. We have found that carboxypeptidase M (CPM) is highly expressed in embryonic LPCs, hepatoblasts, while its expression is decreased along with hepatic maturation. Consistently, CPM expression was transiently induced during hepatic specification from human-induced pluripotent stem cells (hiPSCs). CPM(+) cells isolated from differentiated hiPSCs at the immature hepatocyte stage proliferated extensively in vitro and expressed a set of genes that were typical of hepatoblasts. Moreover, the CPM(+) cells exhibited a mature hepatocyte phenotype after induction of hepatic maturation and also underwent cholangiocytic differentiation in a three-dimensional culture system. These results indicated that hiPSC-derived CPM(+) cells share the characteristics of LPCs, with the potential to proliferate and differentiate bi-directionally. Thus, CPM is a useful marker for isolating hiPSC-derived LPCs, which allows development of a large-scale culture system for producing hepatocytes and cholangiocytes.
    Stem Cell Reports 09/2015; 5(4). DOI:10.1016/j.stemcr.2015.08.008 · 5.37 Impact Factor
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    • "However, the methodology for maintaining HBCs differentiated from hPSCs has not been well investigated. Zhao et al. (2009) have reported that hESC-derived hepatoblastlike cells (sorted N-cadherin-positive cells were used) could be maintained on STO feeder cells. Although a culture system using STO feeder cells for the maintenance of hepatoblast-like cells might be useful, there are two problems. "
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    • "Most of the reagents used for cell culture were obtained from Gibco. Diff-hiPSCs were obtained using a previously described procedure for inducing hepatic progenitors (Zhao et al. 2009). A5 hiPSCs were treated with 10 ng/ml of Activin A and were further cultured with HGF-containing hepatic growth medium. "
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    ABSTRACT: DNA cytosine methylation (5mC) is indispensable for a number of cellular processes, including retrotransposon silencing, genomic imprinting, and X chromosome inactivation in mammalian development. Recent studies have focused on 5-hydroxymethylcytosine (5hmC), a new epigenetic mark or intermediate in the DNA demethylation pathway. However, 5hmC itself has no role in pluripotency maintenance in mouse embryonic stem cells (ESCs) lacking Dnmt1, 3a, and 3b. Here, we demonstrated that 5hmC accumulated on euchromatic chromosomal bands that were marked with di- and tri-methylated histone H3 at lysine 4 (H3K4me2/3) in mouse ESCs. By contrast, heterochromatin enriched with H3K9me3, including mouse chromosomal G-bands, pericentric repeats, human satellite 2 and 3, and inactive X chromosomes, was not enriched with 5hmC. Therefore, enzymes that hydroxylate the methyl group of 5mC belonging to the Tet family might be excluded from inactive chromatin, which may restrict 5mC to 5hmC conversion in euchromatin to prevent nonselective de novo DNA methylation. Electronic supplementary material The online version of this article (doi:10.1007/s10577-012-9317-9) contains supplementary material, which is available to authorized users.
    Chromosome Research 10/2012; 20(7). DOI:10.1007/s10577-012-9317-9 · 2.48 Impact Factor
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