Simple and Quick Method to Obtain a Decellularized, Functional Liver Bioscaffold.

Area of Science:

• Regenerative Medicine
• Tissue Engineering
• Organoid Technology

Background:

• Organ transplantation is critical, driving innovation in organ engineering.
• Organ engineering uses acellular matrices for cell reseeding, but is technically challenging.
• Existing methods often require specific cell isolation and costly reagents.

Purpose of the Study:

• To develop a simplified, cost-effective method for deriving functional in vitro hepatic organoids.
• To create a robust protocol for decellularization suitable for organoid generation.
• To establish a reproducible technique for hepatic organoid production for research applications.

Main Methods:

• A novel protocol for hepatic organoid derivation was established, bypassing the need for parenchymal and non-parenchymal cell selection.
• A trypsin-based enzymatic digestion replaced the standard collagenase perfusion for high-yield decellularization.
• A combination of physical and chemical treatments (deionized water, Triton X, and ammonium hydroxide) was employed for decellularization.

Main Results:

• The method successfully produced functional in vitro hepatic organoids without prior cell sorting.
• High-yield decellularization was achieved using a cost-effective trypsin-based digestion.
• The resulting hepatic constructs maintained original architecture and extracellular matrix components while removing all cellular nucleic acids.

Conclusions:

• This fast, simple, and inexpensive procedure yields functional hepatic organoids.
• The developed method offers a promising approach for toxicological and transplantation studies.
• The technique advances organoid technology by simplifying the generation of complex hepatic structures.