Generation of Functional Liver Sinusoidal Endothelial Cells from Human Pluripotent Stem-Cell-Derived Venous Angioblasts

Blair K Gage; Jeff C Liu; Brendan T Innes; Sonya A MacParland; Ian D McGilvray; Gary D Bader; Gordon M Keller

doi:10.1016/j.stem.2020.06.007

Generation of Functional Liver Sinusoidal Endothelial Cells from Human Pluripotent Stem-Cell-Derived Venous Angioblasts

Cell Stem Cell. 2020 Aug 6;27(2):254-269.e9. doi: 10.1016/j.stem.2020.06.007. Epub 2020 Jul 7.

Authors

Blair K Gage¹, Jeff C Liu², Brendan T Innes³, Sonya A MacParland⁴, Ian D McGilvray⁵, Gary D Bader³, Gordon M Keller⁶

Affiliations

¹ McEwen Stem Cell Institute, University Health Network, Toronto, ON M5G1L7, Canada. Electronic address: [email protected].
² The Donnelly Center, University of Toronto, Toronto, ON M5S3E1, Canada.
³ The Donnelly Center, University of Toronto, Toronto, ON M5S3E1, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON M5G1A8, Canada.
⁴ Multi-Organ Transplant Program, Toronto General Hospital Research Institute, Toronto, ON M5G2C4, Canada; Department of Immunology, University of Toronto, Toronto, ON, M5S1A8, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5G1L7, Canada.
⁵ Multi-Organ Transplant Program, Toronto General Hospital Research Institute, Toronto, ON M5G2C4, Canada.
⁶ McEwen Stem Cell Institute, University Health Network, Toronto, ON M5G1L7, Canada; Department of Medical Biophysics, University of Toronto, Toronto, ON M5G1L7, Canada. Electronic address: [email protected].

PMID: 32640183
DOI: 10.1016/j.stem.2020.06.007

Abstract

Liver sinusoidal endothelial cells (LSECs) form a highly specialized microvasculature that plays a critical role in liver function and disease. To better understand this role, we developed a strategy to generate LSECs from human pluripotent stem cells (hPSCs) by first optimizing the specification of arterial and venous angioblasts and derivative endothelial populations. Induction of a LSEC-like fate by hypoxia, cyclic AMP (cAMP) agonism, and transforming growth factor β (TGF-β) inhibition revealed that venous endothelial cells responded more rapidly and robustly than the arterial cells to upregulate LSEC markers and functions in vitro. Upon intrahepatic transplantation in neonates, venous angioblasts engrafted the liver and generated mature, fenestrated LSECs with scavenger functions and molecular profiles of primary human LSECs. When transplanted into the liver of adult mice, angioblasts efficiently gave rise to mature LSECs with robust factor VIII (FVIII) production. Humanization of the murine liver with hPSC-derived LSECs provides a tractable system for studying the biology of this key liver cell type.

Keywords: FVIII; LSEC; VEGF-A; artery; fenestration; hPSC; humanized hepatic vasculature; hypoxia; liver sinusoidal endothelial cell; sinusoid; vein.

Publication types

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

MeSH terms

Animals
Endothelial Cells*
Hepatocytes
Humans
Liver
Mice
Pluripotent Stem Cells*
Transforming Growth Factor beta

Substances

Transforming Growth Factor beta

Grants and funding

FDN159937/CIHR/Canada