Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study

Xiugong Gao; Rong Li; Patrick Cahan; Yang Zhao; Jeffrey J Yourick; Robert L Sprando

doi:10.1186/s13287-020-01914-1

Hepatocyte-like cells derived from human induced pluripotent stem cells using small molecules: implications of a transcriptomic study

Stem Cell Res Ther. 2020 Sep 11;11(1):393. doi: 10.1186/s13287-020-01914-1.

Authors

Xiugong Gao¹, Rong Li², Patrick Cahan³, Yang Zhao², Jeffrey J Yourick², Robert L Sprando²

Affiliations

¹ Division of 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@fda.hhs.gov.
² Division of 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.
³ Department of Biomedical Engineering, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA.

Abstract

Background: Hepatocyte-like cells (HLCs) derived from human induced pluripotent stem cells (iPSCs) hold great promise in toxicological applications as well as in regenerative medicine. Previous efforts on hepatocyte differentiation have mostly relied on the use of growth factors (GFs) to recapitulate developmental signals under in vitro conditions. Recently, the use of small molecules (SMs) has emerged as an attractive tool to induce cell fate transition due to its superiority in terms of both quality and cost. However, HLCs derived using SMs have not been well characterized, especially on the transcriptome level.

Methods: HLCs were differentiated from human iPSCs using a protocol that only involves SMs and characterized by transcriptomic analysis using whole genome microarrays.

Results: HLCs derived using the SM protocol (HLC_SM) displayed specific hepatic marker expression and demonstrated key hepatic functions. Transcriptomic analysis of the SM-driven differentiation defined a hepatocyte differentiation track and characterized the expression of some key marker genes in major stages of hepatocyte differentiation. In addition, HLC_SM were scored with CellNet, a bioinformatics tool quantifying how closely engineered cell populations resemble their target cell type, and compared to primary human hepatocytes (PHHs), adult liver tissue, fetal liver tissue, HLCs differentiated using GFs (HLC_GF), and commercially available HLCs. Similar to HLC_GF, HLC_SM displayed a mixed phenotype of fetal and adult hepatocytes and had relatively low expression of metabolic enzymes, transporters, and nuclear receptors compared to PHHs. Finally, the differentially expressed genes in HLC_SM compared to HLC_GF and to PHHs were analyzed to identify pathways and upstream transcription regulators which could potentially be manipulated to improve the differentiation of HLCs.

Conclusions: Overall, the present study demonstrated the usefulness of the SM-based hepatocyte differentiation method, offered new insights into the molecular basis of hepatogenesis and associated gene regulation, and suggested ways for further improvements in hepatocyte differentiation in order to obtain more mature HLCs that could be used in toxicological studies.

Keywords: Hepatocyte differentiation; Hepatocyte-like cells; Induced pluripotent stem cells; Microarray; Small molecules; Transcriptomics.

Publication types

Research Support, U.S. Gov't, P.H.S.

MeSH terms

Adult
Cell Differentiation
Computational Biology
Hepatocytes
Humans
Induced Pluripotent Stem Cells*
Transcriptome