[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.


CONCLUSIONS
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.
Kang
Yu-Zhan
YZ
Department of hepatobiliary surgery, Southern Medical University Affiliated Zhujiang Hospital, Guangzhou 510282, China.
Wang
Yan
Y
Gao
Yi
Y
D000818:Animals; D005109:Extracellular Matrix; D005260:Female; D008099:Liver / Q000166:cytology*; D008297:Male; D017830:Octoxynol; D011169:Portal Vein; D051381:Rats; D017207:Rats, Sprague-Dawley; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
D017830:Octoxynol