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A transcriptomic study suggesting human iPSC-derived hepatocytes potentially offer a better in vitro model of hepatotoxicity than most hepatoma cell lines

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Abstract

Hepatocytes derived from human induced pluripotent stem cells (iPSCs) hold great promise as an in vitro liver model by virtue of their unlimited long-term supply, stability and consistency in functionality, and affordability of donor diversity. However, the suitability of iPSC-derived hepatocytes (iPSC-Heps) for toxicology studies has not been fully validated. In the current study, we characterized global gene expression profiles of iPSC-Heps in comparison to those of primary human hepatocytes (PHHs) and several human hepatoma cell lines (HepaRG, HuH-7, HepG2, and HepG2/C3A). Furthermore, genes associated with hepatotoxicity, drug-metabolizing enzymes, transporters, and nuclear receptors were extracted for more detailed comparisons. Our results showed that iPSC-Heps correlate more closely to PHHs than hepatoma cell lines, suggesting that iPSC-Heps had a relatively mature hepatic phenotype that more closely resembles that of adult hepatocytes. HepaRG was the sole exception but nonetheless suffers from lack of donor diversity and poor prediction of hepatotoxicity. The effects of sex differences and DMSO treatment on gene expression of the cellular models were also investigated. Overall, the results presented in the current study suggest that iPSC-Heps represent a reproducible source of human hepatocytes and a promising in vitro model for hepatotoxicity evaluation. Further studies are needed to develop a robust protocol for hepatocyte differentiation towards a more mature adult phenotype.

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Abbreviations

ANOVA:

Analysis of variance

cDNA:

Complimentary deoxyribonucleic acid

cRNA:

Complimentary ribonucleic acid

CYP:

Cytochrome P450

DEG:

Differentially expressed gene

DMEM:

Dulbecco’s Modified Eagle Medium

DMSO:

Dimethyl sulfoxide

FC:

Fold change

FDR:

False discovery rate

HCA:

Hierarchical clustering analysis

HEPES :

4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid

iPSC:

Induced pluripotent stem cell

iPSC-Hep:

Induced pluripotent stem cell-derived hepatocyte

IVT:

In vitro transcription

MEM:

Minimal essential medium

PCA:

Principal component analysis

PHH:

Primary human hepatocyte

RIN:

RNA integrity number

RMA:

Robust multi-array average (algorithm)

RPMI:

Roswell Park Memorial Institute (medium)

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Acknowledgement

The authors thank Drs. Thomas J. Flynn and Menghang Xia for providing primary human hepatocytes and HepG2 cells, respectively, and Dr. Michael F. Santillo for critical review of the manuscript. The authors also thank Drs. Jeffrey J. Yourick, Paddy L. Wiesenfeld, and Robert L. Sprando for their support and guidance on this work. The findings and conclusions presented in this article are those of the authors and do not necessarily represent views, opinions, or policies of the U.S. Food and Drug Administration.

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  1. Division of Applied Regulatory Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, Laurel, MD, 20708, USA

    Xiugong Gao & Yitong Liu

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  1. Xiugong Gao
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  2. Yitong Liu
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Correspondence to Xiugong Gao or Yitong Liu.

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Xiugong Gao and Yitong Liu contributed equally to the study.

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Gao, X., Liu, Y. A transcriptomic study suggesting human iPSC-derived hepatocytes potentially offer a better in vitro model of hepatotoxicity than most hepatoma cell lines. Cell Biol Toxicol 33, 407–421 (2017). https://doi.org/10.1007/s10565-017-9383-z

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  • DOI: https://doi.org/10.1007/s10565-017-9383-z

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