title: The dynamic three-dimensional culture of islet-like clusters in decellularized liver scaffolds.
abstract: Diabetes mellitus is a worldwide metabolic disease which constitutes a major threat to human health. Stem cells with the ability to differentiate into insulin-producing cells (IPCs) could provide unlimited sources of transplanted cells and solve allogeneic rejection problems. The decellularized scaffolds could provide IPCs with tissue microarchitecture and intact vascular systems. The goal of this study was to engineer intact whole rat liver scaffolds and repopulate the stem cell-derived IPCs into the scaffolds to discover whether the decellularized scaffolds could facilitate the growth and development of IPCs. Decellularized liver scaffolds were obtained using 1 % Triton X-100 with 0.1 % ammonium hydroxide. Architecture and composition of the original extracellular matrix were confirmed by morphologic, histological and immunolabeling examinations. Islet-like clusters were differentiated from Wharton's jelly mesenchymal stem cells (WJMSCs) by a three-step induction procedure. The differentiation was evaluated by morphology, RT-PCR, immunofluorescence and glucose stimulation experiments. The islet-like clusters were recellularized into the decellularized scaffolds by the portal-vein infusion method and cultured by the dynamic circulation perfusion device. After cultivation, hematoxylin-eosin staining, immunofluorescence and RT-PCR were conducted. Our results demonstrated that the decellularized rat liver scaffolds have favorable biochemical properties and could support the survival of WJMSC-derived IPCs. In addition, the three-dimensional decellularized scaffolds could enhance the expression of the insulin gene compared with two-dimensional plate culture. In conclusion, these findings suggested that the decellularized scaffolds could provide a suitable platform for cellular activities of IPCs such as survival, differentiation, proliferation and insulin secretion. This study provides fundamental support for regenerating insulin-secreting organs from the decellularized scaffolds combined with stem cell-derived IPCs as a potential clinical application.
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China.|Department of Emergency Surgery, Affiliated Hospital of Nantong University, Nantong, China.
authors.forename: Pengcheng
authors.initials: P
authors.lastname: Zhou
authors.affiliation: Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Nantong, China.
authors.forename: Yibing
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China.
authors.forename: Mingyan
authors.initials: M
authors.lastname: Zhu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China.
authors.forename: Xiangjun
authors.initials: X
authors.lastname: Fan
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China.
authors.forename: Shajun
authors.initials: S
authors.lastname: Zhu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China.
authors.forename: Tianxing
authors.initials: T
authors.lastname: Xu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China.
authors.forename: Di
authors.initials: D
authors.lastname: Wu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China. lyh76@126.com.|Surgical Comprehensive Laboratory, Affiliated Hospital of Nantong University, Nantong, China. lyh76@126.com.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, China. wzw3639@163.com.
authors.forename: Zhiwei
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000818:Animals; D002449:Cell Aggregation / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation; D048430:Cell Shape; D004247:DNA / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D021621:Imaging, Three-Dimensional; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology*; D008099:Liver / Q000166:cytology* / Q000648:ultrastructure; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects; D010477:Perfusion; D010641:Phenotype; D017207:Rats, Sprague-Dawley; D046509:Tissue Culture Techniques / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry*; D059631:Wharton Jelly / Q000166:cytology
chemical_list: D007328:Insulin; D004247:DNA; D005947:Glucose
keywords: Decellularized scaffolds; Diabetes mellitus; Dynamic three-dimensional culture; Islets; Mesenchymal stem cells