[Decellularization technology application in whole liver reconstruct biological scaffold].

OBJECTIVE

To explore an innovative method for preparation of a whole-liver reconstruct scaffold with intact three-dimensional geometry, vasculature and bile duct by decellularization technology.

METHODS

The portal vein was annulated and perfused sequentially with 1%Triton X-100 and 1%SDS for about 4 h, and then was perfused with phosphate buffered saline to dilute SDS residue. The retained structure was evaluated by histological analyses, including macroscopic, Hematoxylin-Eosin staining, Masson's trichrome staining, orcein staining and SEM. The liquid polymer preparation 8% - 10%, which was made of chlorinated poly vinyl chloride (CPVC as solute), acetone (as solvent) and pigment, was injected into portal vein and bile duct to demonstrate the integrity of the portal vein and bile duct. The scaffold was cut into slices with the thickness of about 50 microm and cocultured with C3A cell line.

RESULTS

Macroscopic examination showed that the decellularized liver was transparent and intrahepatic Glisson's system could be observed. H&E staining of slices of decellularized liver demonstrated no intact cells or nuclei existed. Masson trichrome staining revealed collagen retained. Orcein staining showed that there were elastic fibers. SEM showed the network of ECM was intact. C3A-to-scaffold co-culture revealed the scaffold of good biocompatibility.

CONCLUSION

Perfusion with detergents through portal vein for liver decellularization was an efficient method to obtain a completely whole-liver scaffold which can be used for hepatic organ reconstruction.