诱导型多能干细胞体外向胰岛素分泌细胞分化及对血糖的调控  

Insulin-secreting cells from induced pluripotent stem cells regulate blood glucose levels in vitro

作　　者：雷蕾 梁英子 苏映军 马显杰 崔鑫 郭树忠 

机构地区：[1]解放军第四军医大学第一附属医院整形外科,陕西省西安市710032

出　　处：《中国组织工程研究》2016年第28期4210-4217,共8页Chinese Journal of Tissue Engineering Research

基　　金：西京医院转化医学研究项目(XJZT13z03)~~

摘　　要：背景:诱导小鼠多潜能性干细胞定向分化为胰岛素分泌细胞能有效改善糖尿病小鼠血糖水平。目的:在体外将诱导型多能干细胞定向分化的胰岛样细胞团进行相关分子及蛋白水平检测,并观察其在体外和体内胰岛素分泌情况。方法:将体外培养的小鼠诱导型多能干细胞采用联合诱导剂分3个阶段对其进行诱导分化,并对每个诱导阶段形成的内胚层细胞、胰岛前体细胞和成熟胰岛细胞进行形态学观察和PCR检测相关基因的表达水平。对成熟的胰岛样细胞团进行双硫腙染色,ELISA检测体外释放胰岛素功能。将获得的胰岛素分泌细胞移植到糖尿病小鼠肾包膜下,观察其血糖水平变化。结果与结论:(1)体外成功获得的小鼠胰岛样细胞团,其结构呈球形,双硫腙染色呈铁红色,PCR检测到在m RNA水平上表达胰岛素基因;(2)ELISA检测到胰岛样细胞团在受到葡萄糖刺激时能够分泌胰岛素;(3)通过肾包膜方法将细胞团移植到1型糖尿病小鼠体内,对小鼠高血糖有明显的调控作用。BACKGROUND：Mouse pluripotent stem cel s are induced to differentiate into insulin-secreting cel s that can effectively improve blood glucose levels in diabetic mice. OBJECTIVE：To detect mRNA and protein levels of insulin-like cel clusters from induced pluripotent stem cel s and to investigate the function of insulin-secreting cel s in vitro and in vivo. METHODS：Mouse induced pluripotent stem cel s cultured in vitro were induced to differentiate into insulin-secreting cel s using combined inducers through three stages. The morphology of endodermal cel s, islet-derived progenitor cel s and mature islet cel s in each stage was observed and relative gene expression levels were detected by PCR. Mature insulin-like cel clusters underwent dithizone staining and functions of insulin released in vitro were observed by ELISA assay. Final y, the insulin-secreting cel s were transplanted into the subrenal capsule of diabetic mice, and then blood glucose levels were observed. RESULTS AND CONCLUSION：The mature spherical insulin-like cel clusters were successful y obtained in vitro, which were in iron red by dithizone staining, and expression of insulin mRNA was determined by PCR. The insulin-like cel clusters could secrete insulin in response to various blood glucose levels by ELISA assay. In addition, after the cel s clusters were transplanted into the subrenal capsule of mice with type 1 diabetes, the blood glucose levels were marbedly improved.

关 键 词：多潜能干细胞 糖尿病 1型 胰岛素分泌细胞 细胞分化 血糖 组织工程 干细胞 分化 定向诱导分化 胰岛样细胞团 1型糖尿病 胰岛素 内胚层细胞 胰岛前体细胞 成熟胰岛细胞 肾包膜移植 血糖水平 

分 类 号：R394.2[医药卫生—医学遗传学]