Efficient Generation of Functional Hepatocytes From Human Embryonic Stem Cells and Induced Pluripotent Stem Cells by HNF4α Transduction

Abstract

Hepatocyte-like cells from human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) are expected to be a useful source of cells drug discovery. Although we recently reported that hepatic commitment is promoted by transduction of SOX17 and HEX into human ESC- and iPSC-derived cells, these hepatocyte-like cells were not sufficiently mature for drug screening. To promote hepatic maturation, we utilized transduction of the hepatocyte nuclear factor 4α (HNF4α) gene, which is known as a master regulator of liver-specific gene expression. Adenovirus vector-mediated overexpression of HNF4α in hepatoblasts induced by SOX17 and HEX transduction led to upregulation of epithelial and mature hepatic markers such as cytochrome P450 (CYP) enzymes, and promoted hepatic maturation by activating the mesenchymal-to-epithelial transition (MET). Thus HNF4α might play an important role in the hepatic differentiation from human ESC-derived hepatoblasts by activating the MET. Furthermore, the hepatocyte like-cells could catalyze the toxication of several compounds. Our method would be a valuable tool for the efficient generation of functional hepatocytes derived from human ESCs and iPSCs, and the hepatocyte-like cells could be used for predicting drug toxicity.

Sustained Delivery Growth Factors with Polyethyleneimine‐Modified Nanoparticles Promote Embryonic Stem Cells Differentiation and Liver Regeneration

“…The differentiated cells displayed a pronounced capacity to take up ICG in the GF‐PEI‐MSN complex‐treated group (Figure 4B) compared with those of other groups. We also examined the capacity for Dil‐ac‐LDL uptake, which is a critical hepatocyte function 48. As shown in Figure 4C, the positive immunofluorescent signals for Dil‐ac‐LDL uptake in the GF‐PEI‐MSN complex group increased significantly in the hepatic lineage differentiated cells at day 18 compared with those of the control and other treatments.…”

Prediction of interindividual differences in hepatic functions and drug sensitivity by using human iPS-derived hepatocytes

“…Human induced pluripotent stem cell (iPSC)-derived hepatocyte-like cells (HLCs) are expected to be useful for the prediction of DILI in the early phase of drug development. Many groups, including our own, have reported that the human iPS-HLCs have the ability to metabolize drugs, and thus these cells could be used to detect the cytotoxicity of drugs that are known to cause DILI (1,2). However, to accurately predict DILI, it will be necessary to establish a panel of human iPS-HLCs that better represents the genetic variation of the human population because there are large interindividual differences in the drug metabolism capacity and drug responsiveness of hepatocytes (3).…”

Differentiation of odontoblast-like cells from mouse induced pluripotent stem cells by Pax9 and Bmp4 transfection

“…However, these methods still have problems, such as usage of other cell types, the heterogeneity of differentiated cells, and tumorigenicity. Takayama et al [10] succeeded in inducing differentiation of human iPS and embryonic stem cells into nearly homogenous functional hepatocytes by transfection of critical transcription factors for hepatocyte differentiation. Their report indicated that transfection of differentiation-associated genes is a powerful tool to induce differentiation of iPS cells.…”