학술논문
Liver sinusoidal endothelial cells promote the differentiation and survival of mouse vascularised hepatobiliary organoids.
Document Type
Academic Journal
Author
Yap KK; O'Brien Institute, Department of St Vincent's Institute, Victoria, Australia; University of Melbourne Department of Surgery, St Vincent's Hospital Melbourne, Victoria, Australia. Electronic address: kyap@svi.edu.au.; Gerrand YW; O'Brien Institute, Department of St Vincent's Institute, Victoria, Australia.; Dingle AM; O'Brien Institute, Department of St Vincent's Institute, Victoria, Australia.; Yeoh GC; Harry Perkins Institute of Medical Research & Centre for Medical Research, University of Western Australia, Western Australia, Australia.; Morrison WA; O'Brien Institute, Department of St Vincent's Institute, Victoria, Australia; University of Melbourne Department of Surgery, St Vincent's Hospital Melbourne, Victoria, Australia; Australian Catholic University, Victoria, Australia.; Mitchell GM; O'Brien Institute, Department of St Vincent's Institute, Victoria, Australia; University of Melbourne Department of Surgery, St Vincent's Hospital Melbourne, Victoria, Australia; Australian Catholic University, Victoria, Australia.
Source
Publisher: Elsevier Science Country of Publication: Netherlands NLM ID: 8100316 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1878-5905 (Electronic) Linking ISSN: 01429612 NLM ISO Abbreviation: Biomaterials Subsets: MEDLINE
Subject
Language
English
Abstract
The structural and physiological complexity of currently available liver organoids is limited, thereby reducing their relevance for drug studies, disease modelling, and regenerative therapy. In this study we combined mouse liver progenitor cells (LPCs) with mouse liver sinusoidal endothelial cells (LSECs) to generate hepatobiliary organoids with liver-specific vasculature. Organoids consisting of 5x10 3 cells were created from either LPCs, or a 1:1 combination of LPC/LSECs. LPC organoids demonstrated mild hepatobiliary differentiation in vitro with minimal morphological change; in contrast LPC/LSEC organoids developed clusters of polygonal hepatocyte-like cells and biliary ducts over a 7 day period. Hepatic (albumin, CPS1, CYP3A11) and biliary (GGT1) genes were significantly upregulated in LPC/LSEC organoids compared to LPC organoids over 7 days, as was albumin secretion. LPC/LSEC organoids also had significantly higher in vitro viability compared to LPC organoids. LPC and LPC/LSEC organoids were transplanted into vascularised chambers created in Fah -/- /Rag2 -/- /Il2rg -/- mice (50 LPC organoids, containing 2.5x10 5 LPCs, and 100 LPC/LSEC organoids, containing 2.5x10 5 LPCs). At 2 weeks, minimal LPCs survived in chambers with LPC organoids, but robust hepatobiliary ductular tissue was present in LPC/LSEC organoids. Morphometric analysis demonstrated a 115-fold increase in HNF4α+ cells in LPC/LSEC organoid chambers (17.26 ± 4.34 cells/mm 2 vs 0.15 ± 0.15 cells/mm 2 , p = 0.018), and 42-fold increase in Sox9+ cells in LPC/LSEC organoid chambers (28.29 ± 6.05 cells/mm 2 vs 0.67 ± 0.67 cells/mm 2 , p = 0.011). This study presents a novel method to develop vascularised hepatobiliary organoids, with both in vitro and in vivo results confirming that incorporating LSECs with LPCs into organoids significantly increases the differentiation of hepatobiliary tissue within organoids and their survival post-transplantation.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2020 Elsevier Ltd. All rights reserved.)
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2020 Elsevier Ltd. All rights reserved.)