3D microperfusion of mesoscale human microphysiological liver models improves functionality and recapitulates hepatic...

Area of Science:

• Biomedical Engineering
• Hepatology
• Tissue Engineering

Background:

• Accurate in vitro models of the human liver are crucial for drug development and toxicology.
• Existing models struggle to maintain physiological cell densities and replicate liver zonation.
• 3D culture systems are necessary for sustaining liver function and mimicking in vivo conditions.

Purpose of the Study:

• To develop a 3D liver microphysiological system (MPS) that replicates hepatic zonation and functionality.
• To create a platform for long-term culture of primary human hepatocytes (PHH) and hiPSC-derived hepatocytes.
• To investigate zone-specific drug toxicity patterns within the engineered liver model.

Main Methods:

• Fabrication of 3D stereolithography printed hydrogel chips with microvasculature.
• Seeding of PHH and hiPSC-derived hepatocyte-like cells into the chips.
• Perfusion culture and 3D optical oxygen mapping.
• Spatial proteomics analysis to confirm zonation.

Main Results:

• The liver MPS supported long-term viability and improved phenotypes of human liver cells.
• Engineered models recapitulated physiological oxygen gradients, mimicking liver acini.
• Zone-specific acet-ami-no-phen toxicity patterns were observed, correlating with oxygen gradients.
• Spatial proteomics confirmed periportal and perivenous protein distribution.

Conclusions:

• The microperfused liver MPS is a promising platform for phenotypically relevant liver cell culture.
• The model successfully replicates liver zonation and enables study of zone-specific responses.
• This technology advances in vitro models for drug discovery and toxicology assessments.