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短期或长期胰岛保存的最佳方法:胰岛培养、冷藏保存和冷冻保存的比较。

Optimal method for short-term or long-term islet preservation: comparison of islet culture, cold preservation and cryopreservation.

作者信息

Liu Fei, Tian Wencong, Yang Yinan, Zhang Qiong, Zhu Mengmeng, Yang Liang, Yang Lei, Li Jing, Liu Jie, Wu Ping, Yang Kaichiang, Wang Ximo, Shen Yanna, Qi Zhi

机构信息

Department of Histology and Embryology, School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, 300071, China.

出版信息

J Artif Organs. 2014 Dec;17(4):337-43. doi: 10.1007/s10047-014-0777-x. Epub 2014 Jun 19.

Abstract

Islet preservation plays an important role for the success of islet transplantation. To determine the optimal method for islet preservation, we compared the outcomes of islet culture, cold preservation, and cryopreservation in this study. Isolated rat islets were divided into three groups: 37 °C group (conventional culture at 37 °C in RPMI-1640 medium), 4 °C group (cold preservation at 4 °C in University of Wisconsin (UW) solution), and -80 °C group (cryopreservation at -80 °C with dimethyl sulfoxide (DMSO)). Recovery rate, Calcein-AM/PI double staining, insulin release, mRNA level of hypoxia-inducible factor-1α (HIF-1α), and protein level of Bax in islets were examined after short-term (1 day) or long-term (7 days) preservation. After either short-term or long-term preservation, 4 °C group showed higher recovery rate of the islets number, lower percentage of PI positive area, better insulin release ability, and lower expression levels of HIF-1α and Bax in comparison to the 37 or -80 °C group. Meanwhile, islets in 37 °C group showed better function, and down-regulation of HIF-1α and Bax than those in -80 °C group on day 1; however, worse function of islets, up-regulated HIF-1α and Bax in 37 °C group were observed in comparison to -80 °C group on day 7. These results suggest that cold preservation at 4 °C in UW solution is the optimal method in comparison to the conventional culture at 37 °C or cryopreservation at -80 °C for short-term or long-term islet preservation. Furthermore, the potential mechanism may relate to, at least in part, apoptosis induced by the HIF-1α.

摘要

胰岛保存对于胰岛移植的成功起着重要作用。为了确定胰岛保存的最佳方法,我们在本研究中比较了胰岛培养、冷藏保存和冷冻保存的效果。将分离的大鼠胰岛分为三组:37°C组(在RPMI-1640培养基中于37°C常规培养)、4°C组(在威斯康星大学(UW)溶液中于4°C冷藏保存)和-80°C组(在-80°C用二甲基亚砜(DMSO)冷冻保存)。在短期(1天)或长期(7天)保存后,检测胰岛的回收率、钙黄绿素-AM/碘化丙啶(PI)双染、胰岛素释放、缺氧诱导因子-1α(HIF-1α)的mRNA水平以及Bax的蛋白水平。短期或长期保存后,与37°C或-80°C组相比,4°C组的胰岛数量回收率更高、PI阳性面积百分比更低、胰岛素释放能力更好,且HIF-1α和Bax的表达水平更低。同时,在第1天,37°C组的胰岛功能比-80°C组更好,HIF-1α和Bax的下调程度也更大;然而,在第7天,与-80°C组相比,37°C组的胰岛功能更差,HIF-1α和Bax上调。这些结果表明,与在37°C常规培养或在-80°C冷冻保存相比,在UW溶液中于4°C冷藏保存是短期或长期胰岛保存的最佳方法。此外,潜在机制可能至少部分与HIF-1α诱导的细胞凋亡有关。

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