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

TitleExpansion, in vivo-ex vivo cycling, and genetic manipulation of primary human hepatocytes.
Publication TypeJournal Article
Year of Publication2020
AuthorsMichailidis E, Vercauteren K, Mancio-Silva L, Andrus L, Jahan C, Ricardo-Lax I, Zou C, Kabbani M, Park P, Quirk C, Pyrgaki C, Razooky B, Verhoye L, Zoluthkin I, Lu W-Y, Forbes SJ, Chiriboga L, Theise ND, Herzog RW, Suemizu H, Schneider WM, Shlomai A, Meuleman P, Bhatia SN, Rice CM, de Jong YP
JournalProc Natl Acad Sci U S A
Date Published2020 Jan 21

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 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.

Alternate JournalProc. Natl. Acad. Sci. U.S.A.
PubMed ID31915293
PubMed Central IDPMC6983380
Grant ListR01 HL131093 / HL / NHLBI NIH HHS / United States
R01 AI091707 / AI / NIAID NIH HHS / United States
F32 DK107164 / DK / NIDDK NIH HHS / United States
R01 AA027327 / AA / NIAAA NIH HHS / United States
K08 DK090576 / DK / NIDDK NIH HHS / United States
P30 CA014051 / CA / NCI NIH HHS / United States
R01 DK085713 / DK / NIDDK NIH HHS / United States
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