Basic Study
Copyright ©The Author(s) 2015. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Oct 21, 2015; 21(39): 11118-11126
Published online Oct 21, 2015. doi: 10.3748/wjg.v21.i39.11118
Hepatic differentiation of rat induced pluripotent stem cells in vitro
Chao Sun, Jun-Jie Hu, Qin Pan, Yi Cao, Jian-Gao Fan, Guang-Ming Li
Chao Sun, Qin Pan, Yi Cao, Jian-Gao Fan, Guang-Ming Li, Department of Gastroenterology, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
Jun-Jie Hu, Department of Gastroenterology, Shanghai First People’s Hospital, Shanghai Jiao Tong University, Shanghai 200080, China
Author contributions: Sun C performed the experiments, analyzed the data and drafted the article; Hu JJ performed the experiments; Pan Q and Cao Y analyzed the data; Fan JG supervised the study and revised the article; Li GM conceived and designed the study.
Supported by National Natural Science Foundation of China, No. 81000173, No. 81070344 and No. 81100297.
Institutional review board statement: The study did not involve human or animal subjects.
Institutional animal care and use committee statement: The study did not involve human or animal subjects.
Conflict-of-interest statement: The authors declare no financial conflict of interest.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Guang-Ming Li, MD, PhD, Department of Gastroenterology, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, 1665 Kongjiang Road, Shanghai 200092, China. ligm55@163.com
Telephone: +86-21-25077343 Fax: +86-21-65030840
Received: January 22, 2015
Peer-review started: January 24, 2015
First decision: April 14, 2015
Revised: April 23, 2015
Accepted: August 25, 2015
Article in press: August 25, 2015
Published online: October 21, 2015
Processing time: 269 Days and 18.6 Hours
Abstract

AIM: To show the efficient generation of hepatocyte-like cells (HLCs) differentiated from the induced pluripotent stem cells (iPSCs) of rats.

METHODS: Hepatic differentiation was achieved using a three-step protocol with several growth factors. First, rat iPSCs were differentiated into definitive endoderm cells using Activin A and Wnt3a treatment. Then fibroblast growth factor 4 and bone morphogenetic protein 2 were added to the culture medium and used to induce hepatic differentiation. Finally, hepatocyte growth factor, Oncostatin M and dexamethasone were used for hepatic maturation. The liver-related markers and functions of HLCs were assessed at the gene and protein levels.

RESULTS: After endodermal induction, the differentiated cells expressed endodermal markers forkhead box protein A2 and SRY-box containing gene 17 at the mRNA and protein levels. After 20 d of culture, the iPSCs were differentiated into HLCs. These differentiated cells expressed hepatic markers including α-fetoprotein, albumin CK8, CK18, CK19, and transcription factor HNF-4α. In addition, the cells expressed functional proteins such as α1-antitrypsin, cytochrome P450 1A2 and CYP 3A4. They acted like healthy hepatic cells, storing glycogen and taking up indocyanine green and low-density lipoproteins. Also, the rates of urea synthesis (20 d 1.202 ± 0.080 mg/dL vs 0 d 0.317 ± 0.021 mg/dL, P < 0.01) and albumin secretion (20 d 1.601 ± 0.102 mg/dL vs 0 d 0.313 ± 0.015 mg/dL, P < 0.01) increased significantly as differentiation progressed.

CONCLUSION: Rat iPSCs can differentiate into HLCs rapidly and efficiently. These differentiated cells may be an attractive resource for treatment of end-stage liver disease.

Keywords: Hepatic differentiation; Induced pluripotent stem cells; Hepatocyte-like cells; Rat

Core tip: Induced pluripotent stem cells (iPSCs) can differentiate into many kinds of cells, including hepatocytes. However, the most important animal model for studying human diseases, especially liver diseases, is the rat model. This study is the first to show the efficient generation of hepatocyte-like cells (HLCs) differentiated from the iPSCs of rats. Hepatic differentiation was achieved using a three-step protocol. Rat iPSCs were differentiated into HLCs after 20 d of culture. These differentiated cells expressed hepatic markers and exhibited functional hepatic characteristics. These differentiated cells may be an attractive resource for treatment of liver disease.