Strategies to produce hepatocytes and hepatocyte‐like cells from pluripotent stem cells

Abstract

Human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are a potent source for unlimited production of hepatocytes and hepatocyte-like cells that may replace primary human hepatocytes in a variety of fields including liver cell therapy, liver tissue engineering, manufacturing bioartificial liver, modeling inherited and chronic liver diseases, drug screening and toxicity testing. Human ESCs are able to spontaneously form embryoid bodies, which then spontaneously differentiate to various tissue-specific cell lineages containing a total of 10-30% albumin-producing hepatocytes and hepatocyte-like cells. Enrichment of embryoid bodies with the definitive endoderm, from which hepatocytes arise, yields increasing the final ratio of hepatocyte population up by 50-65%. Current strategies of the directed differentiation of human ESCs (and iPSCs) to hepatocytes that reproduce liver embryogenesis by sequential stimulation of culturing ESCs with tissue-specific growth factors result in achieving the differentiation rate up to 60-80%. In the future, directed differentiation of human ESCs and iPSCs to hepatocytes should be further optimized towards generating homogeneous cultures of hepatocytes in order to avoid expensive procedures of separation and isolation of hepatocytes and hepatocyte-like cells.

A Perspective on Efflux Transport Proteins in the Liver

“…Current strategies to differentiate iPS cells into hepatocytes using growth factors achieve differentiation rates of up to 60–80%. (125) The use of stem cell-derived hepatocytes as a substitute for primary human hepatocytes for studying and predicting hepatobiliary efflux of drugs/metabolites will depend on their comparability to mature hepatocytes with respect to morphology as well as transporter and enzyme expression. Further, improvements in differentiation strategies are necessary to achieve 100% differentiation into hepatocytes and to avoid costly isolation and purification processes.…”

Human induced pluripotent stem cells derived hepatocytes: rising promise for disease modeling, drug development and cell therapy

“…As sharing most common features to human ESC, human iPSC are proved to be a suitable source to regenerate hepatocytes, and a variety of strategies have been developed in the past few years (Busletta et al, 2011;Chistiakov and Chistiakov, 2012). Song et al (2009) described the first successful hepatic differentiation of human iPSC, closely followed by several other reports (Liu et al, 2010;Rashid et al, 2010;Si-Tayeb et al, 2010b;Sullivan et al, 2010).…”

Comparative transcriptomics of hepatic differentiation of human pluripotent stem cells and adult human liver tissue

“…Improvements of the differentiation process of stem cells into hPSC-HEP have been achieved gradually in the last few years; however, methodology to generate fully functional hPSC-HEP is still lacking (9,15,23,25,42). The breakthrough of differentiating stem cells into hPSC-HEP with some activity of drug metabolic enzymes was accomplished by mimicking hepatogenesis in vitro (1,7,18,58).…”