title: Three-dimensional culture of mouse pancreatic islet on a liver-derived perfusion-decellularized bioscaffold for potential clinical application.
abstract: The cutting-edge technology of three-dimensional liver decellularized bioscaffold has a potential to provide a microenvironment that is suitable for the resident cells and even develop a new functional organ. Liver decellularized bioscaffold preserved the native extracellular matrix and three-dimensional architecture in support of the cell culture. The goal of this study was to discover if three-dimensional extracellular matrix derived from mouse liver could facilitate the growth and maintenance of physiological functions of mouse isolated islets. We generated a whole organ liver decellularized bioscaffold which could successfully preserve extracellular matrix proteins and the native vascular channels using 1% Triton X-100/0.1% ammonium protocol. To evaluate the potential of decellularized liver as a scaffold for islets transplantation, the liver decellularized bioscaffold was infused with mouse primary pancreatic islets which were obtained through Collagenase P digestion protocol. Its yield, morphology, and quality were estimated by microscopic analysis, dithizone staining, insulin immunofluorescence and glucose stimulation experiments. Comparing the three-dimensional culture in liver decellularized bioscaffold with the orthodoxy two-dimensional plate culture, hematoxylin-eosin staining, immunohistochemistry, and insulin gene expression were tested. Our results demonstrated that the liver decellularized bioscaffold could support cellular culture and maintenance of cell functions. In contrast with the conventional two-dimensional culture, three-dimensional culture system could give rise to an up-regulated insulin gene expression. These findings demonstrated that the liver bioscaffold by a perfusion-decellularized technique could serve as a platform to support the survival and function of the pancreatic islets in vitro. Meanwhile three-dimensional culture system had a superior role in contrast with the two-dimensional culture. This study advanced the field of regenerative medicine towards the development of a liver decellularized bioscaffold capable of forming a neo-organ and could be used as potential clinical application.
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Tianxin
authors.initials: T
authors.lastname: Xu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Mingyan
authors.initials: M
authors.lastname: Zhu
authors.affiliation: Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Yibing
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Di
authors.initials: D
authors.lastname: Wu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Xiangjun
authors.initials: X
authors.lastname: Fan
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Shajun
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Changchun
authors.initials: C
authors.lastname: Lin
authors.affiliation: Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Li
authors.affiliation: Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Lu
authors.affiliation: Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Pengcheng
authors.initials: P
authors.lastname: Zhou
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Jiangsu, People's Republic of China wzw3639@163.com.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000737:chemistry*; D007515:Islets of Langerhans / Q000166:cytology*; D016381:Islets of Langerhans Transplantation; D008099:Liver / Q000737:chemistry*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D061251:Primary Cell Culture / Q000379:methods*; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds / Q000737:chemistry*
keywords: Diabetes mellitus; islets; liver decellularized bioscaffold; three-dimensional culture; transplantation