人脐带间充质干细胞条件培养基对缺氧胰岛的保护作用  

Protective Effect of Conditioned Medium from Human Umbilical Cord-derived Mesenchymal Stem Cells on NICCs Exposed to Hypoxia

作　　者：伍君子 马小倩[1] 杨策军[1] 容鹏飞[1] 聂唯[1] 王维[1] WU Jun-zi;MA Xiao-qian;YANG Ce-jun;RONG Peng-fei;NIE Wei;WANG Wei(The Center of Cell Transplantation and Gene Therapy,the Third Xiangya Hospital,Central South University,Changsha 410013,Hunan,China)

机构地区：[1]中南大学湘雅三医院细胞移植与基因治疗中心,中国湖南长沙410013

出　　处：《生命科学研究》2018年第6期461-467,共7页Life Science Research

基　　金：国家自然科学基金资助项目(81471715);2016年湖南省自然科学基金资助项目(2016JJ3175);2017年中南大学研究生自主探索创新项目(2017zzts891)

摘　　要：为了探讨人脐带间充质干细胞(human umbilical cord-derived mesenchymal stem cells, hUC-MSCs)通过旁分泌作用对缺氧胰岛的保护,将新生猪胰岛细胞团(neonatal porcine islet cell clusters, NICCs)分别用人脐带间充质干细胞条件培养基(human umbilical cord-derived MSC-conditioned medium, hu-MSC-CM)及常规RPMI1640培养基(对照组)置于20%O_2和1%O_2的环境中培养。通过荧光激活细胞分类器、Seahorse线粒体应激实验检测不同培养基对缺氧NICCs活性、功能的影响,利用Western-blot检测缺氧时NICCs生存相关蛋白质的表达水平。实验结果显示,缺氧时hu-MSC-CM组中NICCs细胞得率、活性及功能较对照组均有明显改善(P<0.05);实验组中缺氧诱导因子1α(hypoxia-inducible factor 1α, HIF-1α)表达上调,凋亡及自噬相关蛋白质表达下降。实验结果表明间充质干细胞通过旁分泌作用能有效提高缺氧胰岛的活力和功能。To explore the protective effect of human umbilical cord-derived mesenchymal stem cells(hUCMSCs) on neonatal porcine islet cell clusters(NICCs) exposed to hypoxia by paracrine potential, the NICCs in hUC-MSC-conditioned medium(hu-MSC-CM) and RPMI1640 medium(control group) were cultured under 20% O2 or 1% O2 conditions. The viability and function of NICCs in different media were studied by fluorescence activated cell sorter(FACS) and the extracellular flux assay, respectively. The expression of hypoxia survival related proteins was detected by Western-blot. Compared with the control groups, the results showed that the survival ratio, viability, and function were improved significantly in NICCs cultured in huMSC-CM(P<0.05). The expression of hypoxia-inducible factor 1α(HIF-1α) was up-regulated, and meanwhile,the expression of apoptosis and autophagy associated proteins decreased in Western-blot assay. Experimental results showed that MSCs can improve the viability and function via the paracrine effect.

关 键 词：间充质干细胞(MScs) 新生猪胰岛细胞团(NICCs) 缺氧 活性 功能 

分 类 号：Q955[生物学—动物学]