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首页> 外文期刊>Cell and Tissue Research >The dynamic three-dimensional culture of islet-like clusters in decellularized liver scaffolds
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The dynamic three-dimensional culture of islet-like clusters in decellularized liver scaffolds

机译：去细胞肝支架中胰岛样簇的动态三维培养

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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.
机译：糖尿病是一种全球性代谢疾病，对人类健康构成重大威胁。具有分化为胰岛素产生细胞（IPC）能力的干细胞可以提供无限量的移植细胞来源，并解决同种异体排斥问题。脱细胞的支架可以为IPC提供组织微结构和完整的血管系统。这项研究的目的是工程改造完整的大鼠全肝支架，并将干细胞衍生的IPC重新填充到支架中，以发现脱细胞的支架是否可以促进IPC的生长和发育。使用1％Triton X-100和0.1％氢氧化铵获得脱细胞的肝支架。最初的细胞外基质的结构和组成通过形态，组织学和免疫标记检查得到证实。通过三步诱导程序，将胰岛样簇与沃顿氏果冻间充质干细胞（WJMSC）区分开。通过形态学，RT-PCR，免疫荧光和葡萄糖刺激实验评估分化。通过门静脉输注法将胰岛样簇重新细胞化为脱细胞的支架，并通过动态循环灌注装置进行培养。培养后，进行苏木精-伊红染色，免疫荧光和RT-PCR。我们的结果表明，脱细胞的大鼠肝支架具有良好的生化特性，可以支持WJMSC衍生的IPC的存活。此外，与二维平板培养相比，三维脱细胞支架可以增强胰岛素基因的表达。总之，这些发现表明，脱细胞支架可以为IPC的细胞活性（如生存，分化，增殖和胰岛素分泌）提供合适的平台。这项研究为脱细胞支架与干细胞衍生的IPC结合再生胰岛素分泌器官提供了潜在的临床支持。

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