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首页> 外文期刊>Artificial Organs >Noble gene transduction into pancreatic beta-cells by singularizing islet cells with low doses of recombinant adenoviral vector.
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Noble gene transduction into pancreatic beta-cells by singularizing islet cells with low doses of recombinant adenoviral vector.

机译：通过将胰岛细胞与低剂量的重组腺病毒载体分离来将贵族基因转导至胰腺β细胞。

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Adenovirus-mediated gene transduction into the intact islets has thus far been limited to the surface cells of islets. We evaluated the efficiency of gene delivery by singularization of islets, followed by self-reorganization into islet-like masses. Adenovirus-mediated gene transduction was performed on dispersed islet cells, obtained by two-step digestion of collagenase and ethylene glycol tetraacetic acid/dispase. Good self-reorganization of islet cells in culture was observed until 120 h in islet cells of a control group, a group with a multiplicity of infection (MOI) of 1, and a group with an MOI of 5, with their sizes of 66.7 +/-14.17, 64.0 +/- 15.14, and 60.8 +/- 23.71 microm, respectively. No significant difference in spontaneous reaggregation capability among the islet cell masses was noticed. However, fragmentation of the reaggregated islet mass was observed in the groups with an MOI of 10 and 50 at 72 and 48 h, respectively. The gene transduction rates at an MOI of 0.5, 1, and 5 into islet cells were 56.1 +/- 1.43, 97.6 +/- 0.92, and 100 +/- 0.00%, respectively. The insulin stimulation indices of the reaggregated islet mass at an MOI of 0.5 and 1 were preserved to the level of a nontransduced islet mass; those at an MOI greater than 5 were significantly low. Efficient adenovirus-mediated gene transduction into islet/beta-cells was achieved by adding a process of dispersion of islets into single cells prior to gene transduction without losing the characteristic ability of islet cells to form a functional islet mass in culture.
机译：迄今为止，腺病毒介导的基因转导至完整胰岛的方法仅限于胰岛的表面细胞。我们通过单个化胰岛，然后自我重组为胰岛样肿块来评估基因传递的效率。腺病毒介导的基因转导是在分散的胰岛细胞上进行的，该细胞是通过胶原酶和乙二醇四乙酸/分散酶的两步消化获得的。在对照组，感染复数（MOI）为1的组和MOI为5的组，其大小为66.7 +的胰岛细胞中，观察到培养的胰岛细胞良好的自我重组，直至120 h。 /-14.17、64.0 +/- 15.14和60.8 +/- 23.71微米。胰岛细胞团之间的自发聚集能力没有显着差异。但是，分别在72和48小时的MOI为10和50的组中观察到了重新聚集的胰岛块的碎片。在MOI为0.5、1和5时，胰岛细胞的基因转导率分别为56.1 +/- 1.43、97.6 +/- 0.92和100 +/- 0.00％。 MOI为0.5和1时，重新聚集的胰岛质量的胰岛素刺激指数保持在非转导的胰岛质量水平; MOI大于5的那些显着较低。通过在基因转导之前添加胰岛分散到单个细胞中的过程，而不会丧失胰岛细胞在培养中形成功能性胰岛团块的特征能力，可以实现腺病毒介导的高效基因导入胰岛/β细胞的转导。

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