Optimizing perfusion-decellularization methods of porcine livers for clinical-scale whole-organ bioengineering.
Aim. To refine the decellularization protocol of whole porcine liver, which holds great promise for liver tissue engineering. Methods. Three decellularization methods for porcine livers (1% sodium dodecyl sulfate (SDS), 1% Triton X-100 + 1% sodium dodecyl sulfate, and 1% sodium deoxycholate + 1% sodium dodecyl sulfate) were studied. The obtained liver scaffolds were processed for histology, residual cellular content analysis, and extracellular matrix (ECM) components evaluation to investigate decellularization efficiency and ECM preservation. Rat primary hepatocytes were seeded into three kinds of scaffold to detect the biocompatibility. Results. The whole liver decellularization was successfully achieved following all three kinds of treatment. SDS combined with Triton had a high efficacy of cellular removal and caused minimal disruption of essential ECM components; it was also the most biocompatible procedure for primary hepatocytes. Conclusion. We have refined a novel, standardized, time-efficient, and reproducible protocol for the decellularization of whole liver which can be further adapted to liver tissue engineering.
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Qiong
0000-0002-7130-7494
Q
Wu
Department of Pathology and Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Ji
0000-0001-8413-3270
J
Bao
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Yong-jie
YJ
Zhou
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Yu-jia
YJ
Wang
Laboratory of Pathology and Department of Liver Surgery, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Zheng-gui
ZG
Du
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Yu-jun
YJ
Shi
Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Li
L
Li
Department of Pathology and Laboratory of Pathology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, China.
Hong
H
Bu
D000818:Animals; D005109:Extracellular Matrix / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology; D008099:Liver / Q000166:cytology*; D009924:Organ Culture Techniques / Q000379:methods; D010477:Perfusion; D051381:Rats; D013552:Swine; D023822:Tissue Engineering; D054457:Tissue Scaffolds