Generation of multilineage liver organoids with luminal vasculature and bile ducts from human pluripotent stem cells via modulation of Notch signaling

Introduction PrimeSurface Research Papers

Generation of multilineage liver organoids with luminal vasculature and bile ducts from human pluripotent stem cells via modulation of Notch signaling

Hyo Jin Kim, Gyeongmin Kim, Kyun Yoo Chi, Hyemin Kim, Yu Jin Jang, Seongyea Jo, Jihun Lee, Youngseok Lee,
Dong-Hun Woo, Choongseong Han, Sang Kyum Kim, Han-Jin Park & Jong-Hoon Kim

Stem Cell Res Ther 14, 19 (2023). https://doi.org/10.1186/s13287-023-03235-5

Copyright © Authors 2023
This article is licensed under a Creative Commons Attribution 4.0 International License (CC BY).

Background

The liver is a complex organ composed of hepatocytes and other cells. Hepatocytes (parenchymal cells) perform the majority of functions, but non-parenchymal cells such as hepatic astrocytes and Kupffer cells also play an important role. In vitro modeling of liver disease is important to reproduce the interaction between parenchymal and non-parenchymal cells. Recently, various types of multilineage liver organoids (mLOs) can be generated from human iPS cells, however, the assembly and concurrent differentiation of multiple cell types in individual mLOs remain to be a major challenge. While, most studies have focused on the vascularization of mLOs in host tissue after transplantation in vivo, relatively little information is available on the in vitro formation of luminal vasculature in mLOs themselves.

Research Achievements

The mLOs with luminal blood vessels and bile ducts were generated by assembling hepatic endoderm, hepatic stellate cell-like cells (HscLCs), and endothelial cells derived entirely from hPSCs using PrimeSurfaceTM 96U plate. The results from this study demonstrated that mLOs can reproduce parenchymal and non-parenchymal cell interactions and suggested that their application can advance the precise modeling of liver diseases in vitro.

【Fig.2A】Schematic protocol for self-aggregation and differentiation of mLOs.

Use of PrimeSurface™ in this study

PrimeSurfaceTM 96U plate was used to generate mLOs:

Cells	:	To optimize the ratio and the composition of cell types in mLOs (total of 5 × 104 cells), different ratios of HE/EC/HscLC and combinations (HE + EC + HscLC; HE + HscLC; HE only) of cell types were assembled and seeded.
Medium	:	cold multilineage liver organoid differentiation medium (DM)
Culture	:	days 1 and 3, cold DM was added to the organoid culture. Five days after aggregation, mLOs were transferred to ultra-low attachment 24-well plates and further cultured in DM. The medium was replaced every 3 days.
Results	:	presumptive sprouting of EC networks at the periphery of organoids with HscLCs, while a decrease in organoid size in mLO w/o HscLCs.