Highly Efficient Generation of Human Hepatocyte-Like Cells from Induced Pluripotent Stem Cells

Department of Cell Biology, Division of Pediatric Pathology, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
Hepatology (Impact Factor: 11.06). 01/2010; 51(1):297-305. DOI: 10.1002/hep.23354
Source: PubMed


There exists a worldwide shortage of donor livers available for orthotropic liver transplantation and hepatocyte transplantation therapies. In addition to their therapeutic potential, primary human hepatocytes facilitate the study of molecular and genetic aspects of human hepatic disease and development and provide a platform for drug toxicity screens and identification of novel pharmaceuticals with potential to treat a wide array of metabolic diseases. The demand for human hepatocytes, therefore, heavily outweighs their availability. As an alternative to using donor livers as a source of primary hepatocytes, we explored the possibility of generating patient-specific human hepatocytes from induced pluripotent stem (iPS) cells. CONCLUSION: We demonstrate that mouse iPS cells retain full potential for fetal liver development and describe a procedure that facilitates the efficient generation of highly differentiated human hepatocyte-like cells from iPS cells that display key liver functions and can integrate into the hepatic parenchyma in vivo.

    • "iPSCs were induced to differentiate into endoderm by addition of BMP4/FGF2/Activin A for 2 d followed by Activin A for 3 d as described previously (Fig. 1A; Mallanna and Duncan 2013). At this stage, typically 80%–90% of cells express endodermal markers, including GATA4, CXCR4, and SOX17 (D'Amour et al. 2005; McLean et al. 2007; Si-Tayeb et al. 2010b). The endoderm cells were then induced to adopt a hepatic fate by the addition of BMP4 and FGF2 (Mallanna and Duncan 2013) in the presence or absence of 2.5 µM FGFR inhibitor PD161570 or PD173074. "

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    • "Therefore, we propose that the use of human induced pluripotent stem cells (hiPSCs) differentiated into hepatocyte-like cells (HLCs) could provide an adapted cellular environment to conduct such studies. Indeed, it has been shown that hiPSCs generated from skin fibroblasts of an individual carrying mutations in the LDLR gene and differentiated into HLCs were able to recapitulate the features of ADH (Cayo et al., 2012; Si-Tayeb et al., 2010). Here, we used a convenient source of patient-derived somatic "
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    • "Human iPSK3 cells were derived from human foreskin fibroblasts transfected with plasmid DNA encoding reprogramming factors OCT4, NANOG, SOX2 and LIN28 (kindly provided by Dr. Stephen Duncan, Medical College of Wisconsin) [27] [28]. Human iPSK3 cells were maintained in mTeSR serum-free medium (StemCell Technologies, Inc., Vancouver, Canada) or knockout serum replacement (SR) medium supplemented with FGF-2 (40 ng/ mL) on 6-well plates coated with Geltrex (Life Technologies) [29]. "
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