Department of Gastroenterological Surgery, Graduate School of Medicine, Osaka University, Osaka, Japan.
Graduate School of Frontier Biosciences, Osaka University, Osaka, Japan.
Biomaterials. 2018 Apr;160:82-91. doi: 10.1016/j.biomaterials.2018.01.020. Epub 2018 Jan 16.
Tissue engineering of insulin-secreting cells using alternatives to islet transplantation has been fueled by the development of available materials and fabrication techniques. We have established a cell coating technique that enables the cell surface to be coated with extracellular matrix based on the concept of a layer-by-layer (LbL) assembly. The present study evaluated whether this technique is beneficial for fabricating pancreatic β-cell spheroids using a mouse β-cell line. The well-structured and dense spheroids could immediately be constructed by the LbL-coated cells. In the functional analysis, spheroids with the LbL-coated cells had greater insulin secretion ability with increased expression of the insulin and glucose transporter 2 genes versus spheroids with non-coated cells. In addition, we found that the expression of connexin 36, a gap junction molecule, was upregulated by the LbL cell coating. When spheroids with the LbL-coated cells were syngeneically transplanted in diabetic mice, blood glucose levels immediately decreased and glucose sensitivity significantly improved after intraperitoneal glucose stimulation compared to spheroids with non-coated cells. This cell coating technique would be a clinically applicable approach for fabricating pancreatic β-cell spheroids and treating type 1 diabetes mellitus.
使用替代胰岛移植的方法进行胰岛素分泌细胞的组织工程，受到了可用材料和制造技术的发展的推动。我们已经建立了一种细胞涂层技术，该技术基于层层（LbL）组装的概念，可使细胞表面涂覆细胞外基质。本研究评估了该技术是否有益于使用小鼠β细胞系制造胰岛β细胞球体。通过 LbL 涂层细胞可以立即构建出结构良好且密集的球体。在功能分析中，与未涂层细胞的球体相比，具有 LbL 涂层细胞的球体具有更大的胰岛素分泌能力，并且胰岛素和葡萄糖转运蛋白 2 基因的表达增加。此外，我们发现间隙连接分子连接蛋白 36 的表达通过 LbL 细胞涂层而上调。当将具有 LbL 涂层细胞的球体同种异体移植到糖尿病小鼠中时，与未涂层细胞的球体相比，在腹腔内葡萄糖刺激后，血糖水平立即下降，并且葡萄糖敏感性显著改善。这种细胞涂层技术将是制造胰岛β细胞球体和治疗 1 型糖尿病的一种临床适用方法。
