Generation of Human Liver Chimeric Mice and Harvesting of Human Hepatocytes from Mouse Livers

Rui Wei; Chi-Wa Cheng; Wai-In Ho; Kwong-Man Ng; Miguel A Esteban; Hung-Fat Tse

doi:10.1007/978-1-0716-1979-7_25

Generation of Human Liver Chimeric Mice and Harvesting of Human Hepatocytes from Mouse Livers

Methods Mol Biol. 2022:2429:379-390. doi: 10.1007/978-1-0716-1979-7_25.

Authors

Rui Wei^{1

2}, Chi-Wa Cheng^{1

2}, Wai-In Ho^{1

2}, Kwong-Man Ng^{1

2}, Miguel A Esteban^{2

3

4}, Hung-Fat Tse^{5

6

7}

Affiliations

¹ The Cardiology Division, Department of Medicine, Li Ka Shing Faculty of Medicine, Queen Mary Hospital, The University of Hong Kong, Hong Kong, SAR, China.
² Hong Kong-Guangdong Stem Cell and Regenerative Medicine Research Centre, The University of Hong Kong and Guangzhou Institutes of Biomedicine and Health, Hong Kong, SAR, China.
³ Laboratory of Integrative Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China.
⁴ Joint School of Life Sciences, Guangzhou Medical University and Guangzhou Institutes of Biomedicine and Health, Guangzhou, China.
⁵ The Cardiology Division, Department of Medicine, Li Ka Shing Faculty of Medicine, Queen Mary Hospital, The University of Hong Kong, Hong Kong, SAR, China. hftse@hku.hk.
⁶ Hong Kong-Guangdong Stem Cell and Regenerative Medicine Research Centre, The University of Hong Kong and Guangzhou Institutes of Biomedicine and Health, Hong Kong, SAR, China. hftse@hku.hk.
⁷ Department of Medicine, The University of Hong Kong-Shenzhen Hospital, Shenzhen, China. hftse@hku.hk.

PMID: 35507175
DOI: 10.1007/978-1-0716-1979-7_25

Abstract

Primary human hepatocytes (PHHs) are widely used as an in vitro model to evaluate various aspects of human hepatic physiology and pathology. However, PHHs isolated from the human liver have very limited ability for ex vivo expansion in culture. Fah^-/-/Rag2^-/-/Il2rg^-/- (FRG) mice are proven to be an ideal bioincubator for repopulation of PHHs. The human liver chimeric FRG mouse is not only a humanized animal model for disease study and drug screening in vivo, but also a potential source of PHHs for cellular therapy. This chapter describes experimental protocols to generate chimeric FRG mice with humanized liver and to isolate PHHs from human liver chimeric FRG mice. Using these methods, PHHs can be expanded to more than 100-fold for harvesting.

Keywords: Chimeras; Hepatic function; Hepatocyte; Humanized model; Liver regeneration.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Chimera
Disease Models, Animal
Hepatocytes*
Humans
Liver*
Mice