Article

HNF4A is essential for specification of hepatic progenitors from human pluripotent stem cells

Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
Development (Impact Factor: 6.27). 08/2011; 138(19):4143-53. DOI: 10.1242/dev.062547
Source: PubMed

ABSTRACT The availability of pluripotent stem cells offers the possibility of using such cells to model hepatic disease and development. With this in mind, we previously established a protocol that facilitates the differentiation of both human embryonic stem cells and induced pluripotent stem cells into cells that share many characteristics with hepatocytes. The use of highly defined culture conditions and the avoidance of feeder cells or embryoid bodies allowed synchronous and reproducible differentiation to occur. The differentiation towards a hepatocyte-like fate appeared to recapitulate many of the developmental stages normally associated with the formation of hepatocytes in vivo. In the current study, we addressed the feasibility of using human pluripotent stem cells to probe the molecular mechanisms underlying human hepatocyte differentiation. We demonstrate (1) that human embryonic stem cells express a number of mRNAs that characterize each stage in the differentiation process, (2) that gene expression can be efficiently depleted throughout the differentiation time course using shRNAs expressed from lentiviruses and (3) that the nuclear hormone receptor HNF4A is essential for specification of human hepatic progenitor cells by establishing the expression of the network of transcription factors that controls the onset of hepatocyte cell fate.

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Available from: Karim Si-Tayeb, Jul 26, 2015
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    • "HNF4a was previously shown to suppress HCC development by repressing epithelial–mesenchymal transition , thereby maintaining the epithelial identity of hepatocytes [6]. HNF4a orchestrates a transcription factor network that controls hepatocyte differentiation and liver morphogenesis [7]. At later stages, HNF4a expression is pivotal for the establishment of liver architecture, zonation and functional activity. "
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    • "By day 13, end of S2, HNF4α stained cells were observed. HNF4α is believed to be essential for specification of hepatic progenitors from human pluripotent stem cells (DeLaForest et al., 2011). By day 18, end of S3, the addition of hepatocyte growth factor (HGF) to the culture conditions resulted in high levels of expression of AFP, and indicated that the specified cells had committed to a hepatoblast fate. "
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    ABSTRACT: Maturation of induced pluripotent stem cells (hiPSCs) to hepatocyte-like cells (HLCs) has been proposed to address the shortage of human hepatocytes for therapeutic applications. The purpose of this study was to evaluate hiPSCs, HLCs and hepatocytes, all of human origin, in terms of performance metrics of relevance to cell therapies. hiPSCs were differentiated to HLCs in vitro using an established four-stage approach. We observed that hiPSCs had low oxygen consumption and possessed small, immature mitochondria located around the nucleus. With maturation to HLCs, mitochondria showed characteristic changes in morphology, ultrastructure, and gene expression. These changes in mitochondria included elongated morphology, swollen cristae, dense matrices, cytoplasmic migration, increased expression of mitochondrial DNA transcription and replication-related genes, and increased oxygen consumption. Following differentiation, HLCs expressed characteristic hepatocyte proteins including albumin and hepatocyte nuclear factor 4-alpha, and intrinsic functions including cytochrome P450 metabolism. But HLCs also expressed high levels of alpha fetoprotein, suggesting a persistent immature phenotype or inability to turn off early stage genes. Furthermore, the levels of albumin production, urea production, cytochrome P450 activity, and mitochondrial function of HLCs were significantly lower than primary human hepatocytes. CONCLUSION: - hiPSCs offer an unlimited source of human HLCs. However, reduced functionality of HLCs compared to primary human hepatocytes limits their usefulness in clinical practice. Novel techniques are needed to complete differentiation of hiPSCs to mature hepatocytes.
    Stem Cell Research 06/2012; 9(3):196-207. DOI:10.1016/j.scr.2012.06.004 · 3.91 Impact Factor
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    • "HNF4a was previously shown to suppress HCC development by repressing epithelial–mesenchymal transition , thereby maintaining the epithelial identity of hepatocytes [6]. HNF4a orchestrates a transcription factor network that controls hepatocyte differentiation and liver morphogenesis [7]. At later stages, HNF4a expression is pivotal for the establishment of liver architecture, zonation and functional activity. "
    Journal of Hepatology 06/2012; 57(4):910-2. DOI:10.1016/j.jhep.2012.05.017 · 10.40 Impact Factor
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