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核转染人骨髓间充质干细胞的胰岛素分泌潜能。

Potential of nucleofected human MSCs for insulin secretion.

机构信息

Department of Medical Engineering, Yonsei University College of Medicine, Seoul, Korea.

出版信息

J Tissue Eng Regen Med. 2011 Nov;5(10):761-9. doi: 10.1002/term.371. Epub 2010 Dec 29.

Abstract

The goal of this experiment was to generate insulin-producing human mesenchymal stem cells (hMSCs) as a therapeutic source for type I diabetes mellitus, which is caused by insulin deficiency due to the destruction of islet β cells. In various trials for the treatment of type I diabetes, cell-based therapy using adult stem cells is considered to be one of the most useful candidates for the treatment. In this experiment, a non-viral method called nucleofection was used to transfect hMSCs with pEGFP-C2 and furin-cleavable human preproinsulin gene (hPPI) to produce insulin-secreting cells as surrogate β cells. Transfection efficiency was determined using flow cytometry analysis. Expression and production of insulin were tested using RT-PCR and ELISA. The expression, production and maturation of insulin from the genetically engineered hMSCs showed an increase when compared with a non-transfected control group. Insulin expression from hMSCs using nucleofection in this study has shown the potential for type I diabetes therapy. For further study, an evaluation for in vivo experiments and clinical applications must be supplemented.

摘要

本实验旨在生成产生胰岛素的人间质干细胞(hMSC)作为治疗 I 型糖尿病的一种治疗来源,I 型糖尿病是由于胰岛β细胞破坏导致胰岛素缺乏引起的。在 I 型糖尿病的各种治疗试验中,使用成体干细胞的细胞疗法被认为是最有用的治疗候选者之一。在本实验中,使用一种称为电穿孔的非病毒方法将 pEGFP-C2 和可切割的人胰岛素原基因(hPPI)转染到 hMSC 中,以产生分泌胰岛素的细胞作为替代β细胞。使用流式细胞术分析确定转染效率。使用 RT-PCR 和 ELISA 测试胰岛素的表达和产生。与非转染对照组相比,转染的 hMSC 中胰岛素的表达、产生和成熟均有所增加。本研究中使用电穿孔的基因工程 hMSC 表达胰岛素显示出治疗 I 型糖尿病的潜力。为了进一步研究,必须补充体内实验和临床应用的评估。

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