Liver sinusoidal endothelial cells promote the differentiation and survival of mouse vascularised hepatobiliary organoids

Kiryu K Yap; Yi-Wen Gerrand; Aaron M Dingle; George C Yeoh; Wayne A Morrison; Geraldine M Mitchell

doi:10.1016/j.biomaterials.2020.120091

Liver sinusoidal endothelial cells promote the differentiation and survival of mouse vascularised hepatobiliary organoids

Biomaterials. 2020 Aug:251:120091. doi: 10.1016/j.biomaterials.2020.120091. Epub 2020 May 5.

Authors

Kiryu K Yap¹, Yi-Wen Gerrand², Aaron M Dingle², George C Yeoh³, Wayne A Morrison⁴, Geraldine M Mitchell⁴

Affiliations

¹ 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.
² O'Brien Institute, Department of St Vincent's Institute, Victoria, Australia.
³ Harry Perkins Institute of Medical Research & Centre for Medical Research, University of Western Australia, Western Australia, Australia.
⁴ 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.

PMID: 32408048
DOI: 10.1016/j.biomaterials.2020.120091

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 5x10³ 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 2.5x10⁵ LPCs, and 100 LPC/LSEC organoids, containing 2.5x10⁵ 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/mm² vs 0.15 ± 0.15 cells/mm², p = 0.018), and 42-fold increase in Sox9+ cells in LPC/LSEC organoid chambers (28.29 ± 6.05 cells/mm² vs 0.67 ± 0.67 cells/mm², 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.

Keywords: Bio-engineering; Liver disease; Liver organoids; Liver progenitor cells; Liver sinusoidal endothelial cells; Vascularised chambers.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation
Endothelial Cells*
Hepatocytes
Liver
Mice
Organoids*