A Multiple-Cell Microenvironment in a 3-Dimensional System Enhances Direct Cellular Reprogramming Into Hepatic...

Area of Science:

• Regenerative Medicine
• Cell Biology
• Hepatology

Background:

• Primary human hepatocytes are difficult to proliferate and maintain, necessitating alternative sources for regenerative medicine.
• Liver organoids offer a promising solution by providing a renewable source of functional hepatocytes.

Purpose of the Study:

• To develop an efficient method for generating human induced hepatocytes using liver organoids in a 3D system.
• To evaluate the potential of liver organoids for applications in disease modeling, pharmaceuticals, and transplantation.

Main Methods:

• Human hepatocytes were induced from fibroblasts via lentiviral expression of FOXA3, HNF1A, and HNF4A.
• Liver organoids were constructed using induced hepatocytes, endothelial cells, and stem cells in a 3D microenvironment.
• Gene and protein expression of liver-specific markers were analyzed in both 2D and 3D systems.

Main Results:

• Liver organoids significantly upregulated hepatic transcription factors, marker genes, transporter genes, and metabolic enzyme genes.
• Fibroblast-specific gene expression was decreased in liver organoids.
• Comparable levels of liver-specific proteins (ALB, AAT, HNF4A) were observed in the 3D organoid system.

Conclusions:

• Direct reprogramming within 3D multiple-cell microenvironments is more controllable and efficient than 2D systems.
• Liver organoids demonstrate significant potential for disease modeling, drug development, and cell-based therapies.