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Improved Survival and Function of Rat Cryopreserved Islets by Coculture With Sertoli Cells

机译：与支持细胞共培养改善大鼠冷冻胰岛的存活和功能

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摘要

In order to investigate how to improve the function and survival of cryopreserved islets, we cocultured cryopreserved thawed rat islets with rat Sertoli cells. After thawing, the islets were divided into the Sertoli cell coculture group and the control group. Using light and transmission electron microscopes, we examined the morphology of islets and measured their apoptosis index (AI) and insulin release stimulation index (SI). Moreover, we measured apoptosis protein and mRNA by western-blot and reverse transcription polymerase chain reaction and cytokine concentrations in supernatant by ELISA. We examined islet graft survival time in diabetic mice and detected insulin in grafts by immunohistochemistry. We found that the morphology, AI, and SI of the coculture group were all significantly improved. The relative expression levels of cleaved caspase-3 P20, P11, and caspase-7 in the coculture group were lower than those in the control group. Compared with the control group, the expression level of Bax was decreased, but that of Bcl-2 was increased. After transplantation, islet survival in the coculture group was similar to that of fresh islets but longer than that in the control group. These results suggest that coculture with rat Sertoli cells significantly improves the yield and function of rat cryopreserved thawed islets by effectively reducing islet apoptosis.
机译：为了研究如何改善冷冻保存的胰岛的功能和存活，我们将冷冻保存的解冻的大鼠胰岛与大鼠支持细胞共培养。解冻后，将胰岛分为支持细胞共培养组和对照组。使用光镜和透射电子显微镜，我们检查了胰岛的形态，并测量了它们的凋亡指数（AI）和胰岛素释放刺激指数（SI）。此外，我们通过Western印迹和逆转录聚合酶链反应检测细胞凋亡蛋白和mRNA，并通过ELISA测定上清液中的细胞因子浓度。我们检查了糖尿病小鼠中胰岛移植物的存活时间，并通过免疫组织化学检测了移植物中的胰岛素。我们发现，共培养组的形态，AI和SI均得到明显改善。共培养组中裂解的caspase-3 P20，P11和caspase-7的相对表达水平低于对照组。与对照组相比，Bax的表达水平降低，而Bcl-2的表达水平升高。移植后，共培养组的胰岛存活与新鲜胰岛相似，但比对照组更长。这些结果表明，通过有效减少胰岛细胞凋亡，与大鼠支持细胞共培养可显着提高大鼠冷冻保存的解冻胰岛的产量和功能。

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