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Cell Biology and Toxicology

, Volume 33, Issue 4, pp 407–421 | Cite as

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

Original Article

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.

Keywords

Microarray iPSC-derived hepatocytes Primary human hepatocytes Hepatoma cell lines DMSO Sex 

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)

Notes

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.

Compliance with ethical standards

Conflict of interest statement

The authors declare that they have no conflict of interest.

Supplementary material

10565_2017_9383_MOESM1_ESM.xlsx (10 kb)
Supplementary Table 1 (XLSX 9 kb)
10565_2017_9383_MOESM2_ESM.xlsx (25 kb)
Supplementary Table 2 (XLSX 24 kb)
10565_2017_9383_MOESM3_ESM.xlsx (23 kb)
Supplementary Table 3 (XLSX 22 kb)
10565_2017_9383_MOESM4_ESM.xlsx (24 kb)
Supplementary Table 4 (XLSX 24 kb)

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Copyright information

© US Government 2017

Authors and Affiliations

  1. 1.Division of Applied Regulatory Toxicology, Office of Applied Research and Safety Assessment, Center for Food Safety and Applied NutritionU.S. Food and Drug AdministrationLaurelUSA

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