Generation of a Scaffold-Free Three-Dimensional Liver Tissue via a Rapid Cell-to-Cell Click Assembly Process

Area of Science:

• Biotechnology and Biomedical Engineering
• Hepatology and Regenerative Medicine
• Drug Discovery and Toxicology

Background:

• Significant interest exists in developing in vitro liver organ models for fundamental research and drug development.
• Existing two-dimensional (2D) hepatic models face challenges in replicating the complex three-dimensional (3D) microenvironment of the liver.
• Generating rapid, efficient, scaffold-free 3D co-culture liver models with multiple cell types remains a significant obstacle.

Purpose of the Study:

• To develop and employ a novel cell-surface engineering strategy for creating stable, scaffold-free 3D multi-hepatic cell line co-culture tissue models.
• To establish the first three-cell line (hepatocytes, hepatic endothelial cells, hepatic stellate cells) 3D liver tissue model.
• To compare the functional superiority of the developed 3D liver tissue chips against traditional 2D co-culture monolayers.

Main Methods:

• Utilized cell-surface engineering via liposome delivery and fusion to display bio-orthogonal functional groups on cell membranes.
• Induced specific and stable cell-cell contacts to assemble multiple hepatic cell lines into 3D tissues.
• Employed a rapid intercell click ligation process for assembling hepatocytes, hepatic endothelial cells, and hepatic stellate cells.
• Assessed mitochondrial metabolic activity and evaluated drug toxicity to compare 3D and 2D models.

Main Results:

• Successfully generated a novel three-cell line co-culture 3D liver tissue model.
• Demonstrated stable cell-cell contacts and tissue formation through the developed cell-surface engineering strategy.
• The 3D liver tissue chips exhibited superior function compared to 2D co-culture monolayers, as evidenced by metabolic activity and drug toxicity assessments.

Conclusions:

• The developed cell-surface engineering strategy enables the rapid and efficient construction of scaffold-free 3D multi-hepatic cell line co-culture models.
• This novel 3D liver tissue model provides a more physiologically relevant platform for in vitro studies compared to traditional 2D cultures.
• The findings support the potential of this 3D liver model for advancing drug discovery prioritization and toxicity evaluations.