Expansion, in vivo–ex vivo cycling, and genetic manipulation of primary human hepatocytes

Eleftherios Michailidis, Koen Vercauteren, Liliana Mancio-Silva, Linda Andrus, Cyprien Jahan, Inna Ricardo-Lax, Chenhui Zou, Mohammad Kabbani, Paul Park, Corrine Quirk, Christina Pyrgaki, Brandon Razooky, Lieven Verhoye, Irene Zoluthkin, Wei-Yu Lu, Stuart J. Forbes, Luis Chiriboga, Neil D. Theise, Roland W. Herzog, Hiroshi SuemizuWilliam M. Schneider, Amir Shlomai, Philip Meuleman, Sangeeta N. Bhatia, Charles M. Rice, Ype P. de Jong

Research output: Contribution to journalArticlepeer-review

Abstract

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 hepatocyte-targeted gene therapy approaches.

Original languageEnglish
Pages (from-to)1678-1688
Number of pages11
JournalProceedings of the National Academy of Sciences
Volume117
Issue number3
Early online date8 Jan 2020
DOIs
Publication statusPublished - 21 Jan 2020

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