Center for Stem Cell and Gene Therapies Research and Practice, Kocaeli University, Kocaeli, Turkey; Department of Stem Cell, Institute of Health Sciences, Kocaeli University, Kocaeli, Turkey.
Center for Stem Cell and Gene Therapies Research and Practice, Kocaeli University, Kocaeli, Turkey; Department of Stem Cell, Institute of Health Sciences, Kocaeli University, Kocaeli, Turkey; Department of Histology and Embryology, Faculty of Medicine, Kocaeli University, Kocaeli, Turkey.
Tissue Cell. 2023 Jun;82:102110. doi: 10.1016/j.tice.2023.102110. Epub 2023 May 16.
In this study, it was aimed to provide a therapeutic approach for T1DM by encapsulating the pancreatic islets with mesenchymal stem cells and decellularized pancreatic extracellular matrix to support the survival of islets while maintaining their cellular activity.
Pancreatic extracellular matrix was decellularized using different concentrations of detergent series. After the preparation of the protein-based tissue extracellular matrix was shown to be free of cells or any genetic material by molecular, immunofluorescence and histochemical techniques. Following the homogenization of the decellularized pancreatic extracellular matrix and the analysis of its protein composition by LC-MS, the matrix proteins were incorporated with pancreatic islets and rat adipose tissue-derived MSCs (rAT-MSCs) in alginate microcapsules. Glucose-stimulated insulin secretion property of the islet cells in the microbeads was evaluated by insulin ELISA. The gene expression profile of the encapsulated cells was analyzed by Real-Time PCR.
Unlike the protein composition of whole pancreatic tissue, the decellularized pancreas matrix was free of histone proteins or proteins originated from mitochondria. The protein matrix derived from pancreatic tissue was shown to support the growth and maintenance of the islet cells. When compared to the non-encapsulated pancreatic islet, the encapsulated cells demonstrate to be more efficient in terms of insulin expression.
The extracellular pancreatic matrix obtained in this study was directly used as supplementary in the alginate-based microcapsule enhancing the cell survival. The tissue matrix protein and alginate had a synergistic effect on total insulin secretion, which might have the potential to overcome the insulin deficiency. Despite the improvement in the cell viability and the number, the efficiency of the insulin secretion in response to glucose stimulation from the alginate microcapsules did not meet the expectation when compared with the non-encapsulated pancreatic islets.
本研究旨在通过将胰岛包封在间充质干细胞和脱细胞胰腺细胞外基质中来为 T1DM 提供一种治疗方法,以支持胰岛的存活,同时保持其细胞活性。
使用不同浓度的去污剂系列对胰腺细胞外基质进行脱细胞处理。通过分子、免疫荧光和组织化学技术证明制备的基于蛋白质的组织细胞外基质不含细胞或任何遗传物质。脱细胞胰腺细胞外基质均匀化后,通过 LC-MS 分析其蛋白质组成,将基质蛋白与胰岛和大鼠脂肪组织来源的间充质干细胞(rAT-MSCs)一起包裹在藻酸盐微胶囊中。通过胰岛素 ELISA 评估微珠中胰岛细胞的葡萄糖刺激胰岛素分泌特性。通过 Real-Time PCR 分析封装细胞的基因表达谱。
与整个胰腺组织的蛋白质组成不同,脱细胞胰腺基质不含组蛋白或来自线粒体的蛋白质。从胰腺组织中提取的蛋白质基质支持胰岛细胞的生长和维持。与未封装的胰岛相比,封装的细胞在胰岛素表达方面更有效。
本研究获得的胰腺细胞外基质可直接用作藻酸盐基微胶囊中的补充物,以增强细胞存活。组织基质蛋白和藻酸盐对总胰岛素分泌具有协同作用,这可能有潜力克服胰岛素缺乏。尽管细胞活力和数量有所提高,但与未封装的胰岛相比,藻酸盐微胶囊对葡萄糖刺激的胰岛素分泌效率并未达到预期。