人类间充质干细胞来源的外泌体对猪胰岛抗缺氧能力的影响  

Effect of human mesenchymal stem cell-derived exosomes on anti-anoxic ability of porcine islets

作　　者：聂唯[1,2] 马小倩 陈则夷[2] 杨策军 容鹏飞 王维[1,2] Nie Wei;Ma Xiaoqian;Chen Zeyi;Yang Cejun;Rong Pengfei;Wang Wei(Cell Transplantation and Gene Therapy Institute,the Third Xiang Ya Hospital,Central South University,Hunan 410013,Changsha,China;Engineering and Technology Research Center for Xenotransplantation of Human Province,Hunan 410013,Changsha,China)

机构地区：[1]中南大学湘雅三医院细胞移植与基因治疗研究所,湖南长沙410013 [2]湖南省异种移植工程技术中心,湖南长沙410013

出　　处：《实用器官移植电子杂志》2018年第5期368-374,共7页Practical Journal of Organ Transplantation(Electronic Version)

基　　金：国家自然科学基金项目(30900359);湖南省自然科学基金项目(2016JJ3175);湖南省卫生计生委科研计划课题(C2015-12)

摘　　要：目的探讨来自人类脐带源间充质干细胞条件培养基(human umbilical cord-derived MSCconditioned medium,Hu-MSC-CM)中的外泌体能否增加新生猪胰岛细胞(neonatal porcine islet cell clusters,NICCs)在缺氧环境下的生存和功能。方法 NICCs在低氧条件下(1%O_2)分别用含外泌体的条件培养基、去除外泌体的条件培养基和常规培养基进行培养。荧光激活细胞分选仪和细胞外通量测定法分别检测外泌体对细胞活性和功能的影响,实时定量PCR检测NICCs中缺氧耐受相关基因HIF-1α、PDH2及VEGF的表达。结果与常规培养基组和去除外泌体的条件培养基组相比,含外泌体的条件培养基组培养的NICCs其存活率、活性和功能均有所提高[IEQ:7 800±210比5 894±188、4 740±273,P <0.001;β细胞存活率(%):77.2±7.0比68.8±1.8、61.5±5.1,P <0.05]。含外泌体的条件培养基组孵育的NICCs细胞中HIF-1α、PDH2及VEGF基因表达均显著高于另外两组。结论人类脐带源MSC-条件培养基可以减轻NICCs在缺氧过程中导致的功能障碍,外泌体在诱导低氧抵抗方面发挥重要作用。[Abstract] Objective To investigated whether exosoines froln huinan umbilical cord-derived MSC- conditioned medium （hu-MSC-CM） could increase the survival and function of neonatal porcine islet cell clusters （NICCs） exposed to hypoxia. Methods NICCs were cultured with Hu-MSC-CM and native medium RPMI- 1640 （control） under hypoxic conditions （1%O2）, with or without exosoines. The effects of exosoines on NICCs viability and function in vitro were examined by FACS and the extracellular Flux assay, respectively. The expression of HIF-1α, PDH2 and VEGF related with hypoxia tolerance genes in NICCs were detected by real-tiine quantitative PCR. Results Compared with NICCs cultured group with RPMI-1640 medium and Hu-MSC-CM group without exosoines, the survival rate, viability and function were increased in Hu-MSC-CM group with exosoines [ IEQ ： 7 800 ± 210 vs. 5 894 ± 188 4 740 ± 273, P 〈 0.001 ; β cell viability （%） ： 77.2 ± 7.0 vs. 68.8 ± 1.8,61.5 ± 5.1, P 〈 0.05]. The expression of HIF-1α, PDH2 and VEGF in Hu-MSC-CM group with exosoines was significantly higher than the other two groups. Conclusion Hu-MSC-CM could protect NICCs from hypoxia-induced dysfunction, and exosomes played an important role in hypoxic resistance.

关 键 词：间充质干细胞 新生猪胰岛细胞 低氧 功能 活性 

分 类 号：R657.5[医药卫生—外科学]