人脐血间充质干细胞向胰岛素分泌细胞分化的研究  

Study on differentiation of mesenchymal stem cells derived from human umbilical cord blood into insulin secreting cells

作　　者：迟作华[1] 陆琰[1] 张洹[1] 

机构地区：[1]暨南大学医学院血液病研究所,广州510632

出　　处：《中华血液学杂志》2008年第10期679-683,共5页Chinese Journal of Hematology

基　　金：国务院侨办重点学科建设基金（2005）;广东省自然科学基金（06300546）;国家自然科学基金（30670902）

摘　　要：目的探讨脐血间充质干细胞（MSC）向胰岛素分泌细胞分化的潜能及其分化诱导条件。方法分离脐血有核细胞，将其置于Mesencuh^TM培养基中进行培养，并利用贴壁法进行纯化、扩增。扩增后的脐山MSC用表皮生长因子（EGF）、β－巯基乙醇和高糖进行诱导。采用RT—PCR、胰岛素免疫细胞化学染色方法对诱导后的细胞进行鉴定，定量检测胰岛素分泌水平及其对葡萄糖刺激的反应性；移植到链脲佐菌素诱导的糖尿病小鼠体内，观测其往体内的降糖作用。结果诱导后细胞形态发生明显变化，细胞变圆且聚集成团；RT—PCR分析显示，细胞表达部分胰岛相关基因；细胞的胰岛素免疫细胞化学染色为附降；而且细胞能分泌少量[（0．37±0．06）mU／L]胰岛素，并对糖刺激具有反应性，刺激指数为1．76；诱导后的细胞移植能降低糖尿病小鼠的血糖。结论脐血MSC具有向胰岛素分泌细胞分化的潜能。Objective To investigate the differential potential of mesenchymal stern cells（MSCs） derived from human umbilical cord blood （hUCB） into insulin-secreting cells and its inducing condition. Methods UCB nucleated cells （NCs） were isolated and cultured in MesencultTM media. The obtained UCB MSC were purified bv adherence method and expanded. Then they were induced with epidermal growth factor （EGF） , B-mercaptoethanol and high concentration of glucose. The induced cells were identified by RT-PCR. Intracellular insulin was examined by immunocytochemistry. The quantity of insulin secretion and glucosesimulated insulin release were examined by chemiluminescence immunoassay. The induced cells were also transplanted into renal subcapsular space of STZ-induced hyperglycemic mice to observe the in vivo lowering effect on hyperglycemia. Results The induced cells morphologically became round and were gathering into a mass. The expression of some genes related to pancreatic islet was found by RT-PCR. Chemiluminescence immunoassay showed insulin positivity and the cells secreted a low concentration of insulin [ （0.37 ±0. 06） mU/L]. The induced cells responded to high glucose challenge with a stimulation index of 1.76. After those cells grafted into renal subcapsularly there was an in vivo lowering effect on blood glucose level on STZ hyperglycemic miee. Conclusion MSCs from UCB can differentiated into insulin secreting cells.

关 键 词：胎血 间充质干细胞 细胞分化 胰岛素 糖尿病 

分 类 号：R686[医药卫生—骨科学]