Generation of Functional Endodermal Hepatic Organoids

Area of Science:

• Stem cell biology
• Organoid technology
• Hepatology

Background:

• Organoid technology enables in vitro generation of human organ-like mini-structures.
• Advances in organoid models are crucial for disease modeling, developmental biology, and drug discovery.
• Induced pluripotent stem cell (iPSC)-derived liver organoids are key for generating hepatocytes.

Purpose of the Study:

• To present a robust and reproducible protocol for generating hepatic organoids from pluripotent stem cells.
• To establish a fast and efficient method for creating functional liver organoids within two weeks.
• To provide an alternative source for producing functional hepatocytes.

Main Methods:

• Direct differentiation of iPSCs into endodermal cells.
• FACS-enrichment of EpCAM-positive cells.
• Establishment of hepatic organoids using expansion medium to create 3D endoderm-derived hepatic organoids (eHEPOs).

Main Results:

• Hepatic organoids (eHEPOs) were generated within 2 weeks.
• eHEPOs mimic essential hepatocyte functions, including albumin secretion, glycogen storage, and cytochrome P450 activity.
• eHEPOs exhibit liver-specific gene expression, polarized epithelial cells, and bile canaliculi, and can be serially passaged for up to a year without losing differentiation capacity.

Conclusions:

• The developed protocol provides a fast, efficient, and reproducible method for generating functional hepatic organoids (eHEPOs) from pluripotent stem cells.
• eHEPOs serve as a valuable in vitro model for studying liver biology and disease, and for drug discovery.
• Long-term expansion and differentiation capacity of eHEPOs offer a promising alternative source for functional hepatocytes.