Title | Expansion, in vivo-ex vivo cycling, and genetic manipulation of primary human hepatocytes. |
Publication Type | Journal Article |
Year of Publication | 2020 |
Authors | Michailidis 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 |
Journal | Proc Natl Acad Sci U S A |
Volume | 117 |
Issue | 3 |
Pagination | 1678-1688 |
Date Published | 2020 Jan 21 |
ISSN | 1091-6490 |
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 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. |
DOI | 10.1073/pnas.1919035117 |
Alternate Journal | Proc. Natl. Acad. Sci. U.S.A. |
PubMed ID | 31915293 |
PubMed Central ID | PMC6983380 |
Grant List | R01 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 |