Laboratoire de bio-ingénierie et de biophysique de l'Université de Sherbrooke, Department of Chemical and Biotechnological Engineering, Université de Sherbrooke, Sherbrooke, Canada.
Centre de recherche du CHUS, Faculté de médecine et des sciences de la santé, Sherbrooke, Canada.
J Biomed Mater Res B Appl Biomater. 2023 Nov;111(11):1890-1902. doi: 10.1002/jbm.b.35292. Epub 2023 Jun 12.
Loss in the number or function of insulin-producing β-cells in pancreatic islets has been associated with diabetes mellitus. Although islet transplantation can be an alternative treatment, complications such as apoptosis, ischaemia and loss of viability have been reported. The use of decellularized organs as scaffolds in tissue engineering is of interest owing to the unique ultrastructure and composition of the extracellular matrix (ECM) believed to act on tissue regeneration. In this study, a cell culture system has been designed to study the effect of decellularized porcine bladder pieces on INS-1 cells, a cell line secreting insulin in response to glucose stimulation. Porcine bladders were decellularized using two techniques: a detergent-containing and a detergent-free methods. The resulting ECMs were characterized for the removal of both cells and dsDNA. INS-1 cells were not viable on ECM produced using detergent (i.e., sodium dodecyl sulfate). INS-1 cells were visualized following 7 days of culture on detergent-free decellularized bladders using a cell viability and metabolism assay (MTT) and cell proliferation quantified (CyQUANT™ NF Cell Proliferation Assay). Further, glucose-stimulated insulin secretion and immunostaining confirmed that cells were functional in response to glucose stimulation, as well as they expressed insulin and interacted with the detergent-free produced ECM, respectively.
胰岛中胰岛素产生β细胞的数量或功能的丧失与糖尿病有关。尽管胰岛移植可以作为一种替代治疗方法，但已经报道了细胞凋亡、缺血和活力丧失等并发症。由于细胞外基质（ECM）的独特超微结构和组成被认为对组织再生有作用，因此使用去细胞化器官作为组织工程中的支架引起了人们的兴趣。在这项研究中，设计了一种细胞培养系统来研究去细胞化猪膀胱片对 INS-1 细胞的影响，INS-1 细胞是一种对葡萄糖刺激分泌胰岛素的细胞系。使用两种技术对猪膀胱进行去细胞化：一种含去污剂的方法和一种无去污剂的方法。通过去除细胞和 dsDNA 来对所得 ECM 进行表征。在使用去污剂（即十二烷基硫酸钠）制备的 ECM 上，INS-1 细胞无法存活。使用细胞活力和代谢测定法（MTT）和细胞增殖定量（CyQUANT™ NF 细胞增殖测定法），在无去污剂去细胞化膀胱上培养 7 天后，观察到 INS-1 细胞。此外，葡萄糖刺激胰岛素分泌和免疫染色证实细胞对葡萄糖刺激有反应功能，并且它们分别表达胰岛素并与无去污剂制备的 ECM 相互作用。
