Expansion, in vivo–ex vivo cycling, and genetic manipulation of primary human hepatocytes
Name
1678.full.pdf
Description
Published version
Size
1.49 MB
Format
Adobe PDF
Checksum (MD5)
3eb09600506284d8d36c84fa80dbd34a
Author(s)
Michailidis, Eleftherios
Vercauteren, Koen
Mancio-Silva, Liliana
Andrus, Linda
Jahan, Cyprien
Ricardo-Lax, Inna
Zou, Chenhui
Kabbani, Mohammad
Park, Paul
Quirk, Corrine
Date Issued
2020
Journal
Proceedings of the National Academy of Sciences of the United States of America
Publisher
Proceedings of the National Academy of Sciences
Version
Final published version
Abstract
© 2020 National Academy of Sciences. All rights reserved. Primary human hepatocytes (PHHs) are an essential tool for modeling drug metabolism and liver disease. However, variable plating efficiencies, short lifespan in culture, and resistance to genetic manipulation have limited their use. Here, we show that the pyrrolizidine alkaloid retrorsine improves PHH repopulation of chimeric mice on average 10-fold and rescues the ability of even poorly plateable donor hepatocytes to provide cells for subsequent ex vivo cultures. These mouse-passaged (mp) PHH cultures overcome the marked donor-to-donor variability of cryopreserved PHH and remain functional for months as demonstrated by metabolic assays and infection with hepatitis B virus and Plasmodium falciparum. mpPHH can be efficiently genetically modified in culture, mobilized, and then recultured as spheroids or retransplanted to create highly humanized mice that carry a genetically altered hepatocyte graft. Together, these advances provide flexible tools for the study of human liver disease and evaluation of hepatocytetargeted gene therapy approaches.
MIT Department
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Howard Hughes Medical Institute
Koch Institute for Integrative Cancer Research at MIT
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Terms of Use
Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.
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DOI of Published Version
10.1073/PNAS.1919035117
https://doi.org/10.1073/PNAS.1919035117
