Ming Xin Pan1, Peng Yun Hu2, Yuan Cheng1, Li Quan Cai1, Xiao Hui Rao1, Yan Wang3, Yi Gao4. 1. Department of Hepatobiliary Surgery, Southern Medical University, Guangzhou, Guangdong Province, China; Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong Province, China. 2. Department of Tumor Surgery, Xinxiang Central Hospital, Xinxiang, Henan Province, China. 3. Department of Hepatobiliary Surgery, Southern Medical University, Guangzhou, Guangdong Province, China; Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong Province, China. Electronic address: nn1012@gmail.com. 4. Department of Hepatobiliary Surgery, Southern Medical University, Guangzhou, Guangdong Province, China; Institute of Regenerative Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong Province, China. Electronic address: gaoyi6146@163.com.
Abstract
BACKGROUND/ PURPOSE: Using gradient ionic detergent, we optimized the preparation procedure for the decellularized liver biologic scaffold, and analyzed its immunogenicity and biocompatibility. METHODS: EDTA, hypotonic alkaline solution, Triton X-100, and gradient sodium dodecyl sulfate (1%, 0.5%, and 0.1%, respectively) were prepared for continuous perfusion through the hepatic vascular system. The decellularization of the liver tissue was performed with the optimized reagent buffer and washing protocol. In addition, the preservation of the original extracellular matrix was observed. To analyze its biocompatibility, the scaffold was embedded in a heterologous animal and the inflammation features, including the surrounding cell infiltration and changes of the scaffold architecture, were detected. The cell-attachment ability was also validated by the perfusion culture of HepG2 cells with the scaffold. RESULTS: By using gradient ionic detergent, we completed the decellularization process in approximately 5 h, which was shorter than >10 hours in previous experiments (p<0.001). The extracellular matrix was kept relatively intact, with no obvious inflammatory cellular infiltration or structural damage in the grafted tissue. The engraftment efficiencies of HepG2 were 86±5% (n=8). The levels of albumin and urea synthesis were significantly superior to the ones in traditional two-dimensional culture. CONCLUSION: The current new method can be used efficiently for the decellularization of the liver biologic scaffold with satisfying biocomparability for application both in vivo and in vitro.
BACKGROUND/ PURPOSE: Using gradient ionic detergent, we optimized the preparation procedure for the decellularized liver biologic scaffold, and analyzed its immunogenicity and biocompatibility. METHODS:EDTA, hypotonic alkaline solution, Triton X-100, and gradient sodium dodecyl sulfate (1%, 0.5%, and 0.1%, respectively) were prepared for continuous perfusion through the hepatic vascular system. The decellularization of the liver tissue was performed with the optimized reagent buffer and washing protocol. In addition, the preservation of the original extracellular matrix was observed. To analyze its biocompatibility, the scaffold was embedded in a heterologous animal and the inflammation features, including the surrounding cell infiltration and changes of the scaffold architecture, were detected. The cell-attachment ability was also validated by the perfusion culture of HepG2 cells with the scaffold. RESULTS: By using gradient ionic detergent, we completed the decellularization process in approximately 5 h, which was shorter than >10 hours in previous experiments (p<0.001). The extracellular matrix was kept relatively intact, with no obvious inflammatory cellular infiltration or structural damage in the grafted tissue. The engraftment efficiencies of HepG2 were 86±5% (n=8). The levels of albumin and urea synthesis were significantly superior to the ones in traditional two-dimensional culture. CONCLUSION: The current new method can be used efficiently for the decellularization of the liver biologic scaffold with satisfying biocomparability for application both in vivo and in vitro.
Authors: Panagiotis Maghsoudlou; Fanourios Georgiades; Holly Smith; Anna Milan; Panicos Shangaris; Luca Urbani; Stavros P Loukogeorgakis; Benedetta Lombardi; Giuseppe Mazza; Charlotte Hagen; Neil J Sebire; Mark Turmaine; Simon Eaton; Alessandro Olivo; Jasminka Godovac-Zimmermann; Massimo Pinzani; Paul Gissen; Paolo De Coppi Journal: PLoS One Date: 2016-05-09 Impact factor: 3.240
Authors: Giuseppe Mazza; Krista Rombouts; Andrew Rennie Hall; Luca Urbani; Tu Vinh Luong; Walid Al-Akkad; Lisa Longato; David Brown; Panagiotis Maghsoudlou; Amar P Dhillon; Barry Fuller; Brian Davidson; Kevin Moore; Dipok Dhar; Paolo De Coppi; Massimo Malago; Massimo Pinzani Journal: Sci Rep Date: 2015-08-07 Impact factor: 4.379