Basic Study
Copyright ©The Author(s) 2017. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jul 21, 2017; 23(27): 4935-4941
Published online Jul 21, 2017. doi: 10.3748/wjg.v23.i27.4935
Human liver chimeric mouse model based on diphtheria toxin-induced liver injury
Xiao-Nan Ren, Rong-Rong Ren, Hua Yang, Bo-Yin Qin, Xiu-Hua Peng, Li-Xiang Chen, Shun Li, Meng-Jiao Yuan, Chao Wang, Xiao-Hui Zhou
Xiao-Nan Ren, Rong-Rong Ren, Hua Yang, Bo-Yin Qin, Xiu-Hua Peng, Li-Xiang Chen, Shun Li, Meng-Jiao Yuan, Chao Wang, Xiao-Hui Zhou, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China
Xiao-Hui Zhou, Key Laboratory of Medical Molecular Virology, Ministry of Education and Health, Fudan University, Shanghai 201508, China
Author contributions: Ren XN and Zhou XH designed the experiments; Ren XN, Ren RR and Yang H performed the majority of experiments; Ren XN and Zhou XH analyzed the data; Qin BY, Peng XH, Chen LX, Yuan MJ and Wang C contributed to genotyping; Ren XN and Zhou XH wrote the paper; Li S revised the paper.
Supported by Shanghai Science and Technology Development Foundation Project, No. 12140900300; Shanghai Municipal Commission of Health and Family Planning Project, No. 20144Y0073; Shanghai Public Health Clinical Center Project, No. 2014M08; and National Science and Technology Major Project, No. 2017ZX10304402-001-012.
Institutional animal care and use committee statement: All procedures involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of Shanghai Public Health Clinical Center Affiliated to Fudan University.
Conflict-of-interest statement: No conflicts of interest exist in this study.
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: Xiao-Hui Zhou, MD, PhD, Professor, Shanghai Public Health Clinical Center, Fudan University, 2901 Caolang Road, Jin Shan District, Shanghai 201508, China. zhouxiaohui@shaphc.org
Telephone: +86-21-37990333 Fax: +86-21-37990333
Received: January 25, 2017
Peer-review started: February 1, 2017
First decision: March 16, 2017
Revised: April 1, 2017
Accepted: June 1, 2017
Article in press: June 1, 2017
Published online: July 21, 2017
Processing time: 175 Days and 17 Hours
Abstract
AIM

To establish an inducible liver injury mouse model and transplant human hepatocytes to obtain liver-humanized mice.

METHODS

We crossed three mouse strains, including albumin (Alb)-cre transgenic mice, inducible diphtheria toxin receptor (DTR) transgenic mice and severe combined immune deficient (SCID)-beige mice, to create Alb-cre/DTR/SCID-beige (ADSB) mice, which coincidentally harbor Alb-cre and DTR transgenes and are immunodeficient. As the Cre expression is driven by the liver-specific promoter Alb (encoding ALB), the DTR stop signal flanked by two loxP sites can be deleted in the ADSB mice, resulting in DTR expression in the liver. ADSB mice aged 8-10 wk were injected intraperitoneally (i.p.) with diphtheria toxin (DT) and liver damage was assessed by serum alanine aminotransferase (ALT) level. Two days later, mouse livers were sampled for histological analysis, and human hepatocytes were transplanted into the livers on the same day. A human ALB enzyme-linked immunosorbent assay was performed 7, 14, 21 and 28 d after transplantation. Human CD68 immunohistochemistry was performed 30 and 90 d after transplantation.

RESULTS

We crossed Alb-cre with DTR and SCID-beige mice to obtain ADSB mice. These mice were found to have liver damage 4 d after i.p. injection of 2.5 ng/g bodyweight DT. Bodyweight began to decrease on day 2, increased on day 7, and was lowest on day 4 (range, 10.5%-13.4%). Serum ALT activity began to increase on day 2 and reached a peak value of 289.7 ± 16.2 IU/mL on day 4, then returned to background values on day 7. After transplantation of human liver cells, peripheral blood human ALB level was 1580 ± 454.8 ng/mL (range, 750.2-3064.9 ng/mL) after 28 d and Kupffer cells were present in the liver at 30 d in ADSB mice.

CONCLUSION

Human hepatocytes were successfully repopulated in the livers of ADSB mice. The inducible mouse model of humanized liver in ADSB mice may have functional applications, such as hepatocyte transplantation, hepatic regeneration and drug metabolism.

Keywords: Liver disease; Liver injury; Diphtheria toxin; Liver chimeric mouse model

Core tip: We established a novel liver chimeric mouse model following liver damage caused by intraperitoneal injection of diphtheria toxin (DT), and transplanted human hepatocytes to obtain liver-humanized mice. After 28 d, human albumin was detected in these mice. Human hepatocytes were successfully repopulated in the livers of triple-crossed albumin-cre transgenic mice, inducible DT receptor transgenic mice and severe combined immune deficient-beige mice [i.e., Alb-cre/DTR/SCID-beige (ADSB) mice]. Our inducible mouse model of humanized liver in ADSB mice may have functional applications, such as studies on hepatocyte transplantation, hepatic regeneration and drug metabolism.