Matrix-induced liver cell aggregates (MILCA) for bioartificial liver use

Area of Science:

• Biotechnology
• Hepatology
• Tissue Engineering

Background:

• Ex vivo liver cell culture is crucial for bioartificial liver development.
• Existing methods face challenges in maintaining liver microstructure and function.

Purpose of the Study:

• To develop a novel method for creating matrix-induced liver cell aggregates (MILCA) using collagen-coated beads.
• To assess the viability, microstructure, and metabolic function of MILCA.
• To evaluate MILCA performance in a hollow-fiber bioreactor using plasma from patients with fulminant hepatic failure.

Main Methods:

• Porcine hepatocytes were isolated and cultured to form MILCA on collagen-coated beads.
• Cell viability was assessed using trypan blue exclusion and LDH release.
• Cytochrome P-450 activity was measured by 7-hydroxycoumarine formation.
• MILCA ultrastructure was analyzed via transmission electron microscopy.
• MILCA were tested in hollow-fiber bioreactors with patient plasma, monitoring drug clearance and metabolite production.

Main Results:

• MILCA exhibited high viability (>90%) and preserved liver-like ultrastructure, including cell polarity and bile canaliculi.
• Initial Cytochrome P-450 activity was comparable to controls, but decreased over 24 hours compared to monolayer cultures.
• In bioreactor studies, MILCA cleared lidocaine within 3 hours and reduced plasma lactate, bilirubin, and ammonia levels.
• MILCA also increased non-essential amino acid levels in patient plasma.

Conclusions:

• MILCA represent a viable method for generating functional liver cell aggregates with preserved microstructure.
• MILCA demonstrate potential for use in bioartificial liver devices, capable of detoxification and metabolic functions.
• Further research is needed to optimize MILCA for long-term hepatic support and to fully elucidate their therapeutic potential.