胰岛新生相关蛋白肽在促进脐带间充质干细胞分化为胰岛素分泌细胞中的作用  

Role of INGAP-pp in the differentiation of hUCMSCs into insulin producing cells

作　　者：陈桃 王云帅[3] 谷乐辉[1,2] 周淑艳 叶剑[3] 简优强[3] 李富荣[2] 齐晖[2] 

机构地区：[1]暨南大学第二临床医学院(深圳市人民医院)风湿免疫科,广东深圳518020 [2]暨南大学第二临床医学院(深圳市人民医院)干细胞与细胞治疗重点实验室,广东深圳518020 [3]暨南大学第二临床医学院(深圳市人民医院)肝胆外科,广东深圳518020

出　　处：《细胞与分子免疫学杂志》2013年第2期141-145,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81270857);广东省自然科学基金(6027540);深圳市科技计划重点项目(201001005)

摘　　要：目的探索人脐带间充质干细胞(hUCMSC)向胰岛素分泌细胞(IPCs)分化的高效、定向诱导方案。方法在传统诱导方案基础上,加入胰岛新生相关蛋白肽,诱导hUCMSCs分化为IPCs,并通过形态学观察、流式细胞术、荧光定量PCR、免疫荧光、ELISA鉴定IPCs。结果改良组IPCs的C肽阳性细胞率较传统组有明显提高(P<0.05),胰腺发育相关转录因子PDX-1、Ngn3、NeuroD1、MafA、insulin、Glut-2 mRNA表达明显提高(P<0.05)。同时,改良组蛋白C肽、PDX-1、Nkx6.1表达阳性,葡萄糖刺激试验证实胰岛素及C肽分泌能力也明显增强(P<0.05)。结论胰岛新生相关蛋白肽可促进hUCMSCs分化为IPCs,但仍未达到正常人胰岛β细胞功能。Objective To find out a protocol for the efficient and directional differentiation from human umbilical cord mesenchymal stem cells（hUCMSCs） into insulin-producing cells（IPCs）.Methods HUCMSCs were induced to differentiate into IPCs by an improved protocol based on a traditional protocol with addition of islet neogenesis-associated protein-pp（INGAP-pp）,and the IPCs were identified by morphological observation,flow cytometry,real-time quantitative PCR,immunofluorescence and ELISA.Results Compared with the traditional protocol,the improved protocol resulted in the higher rate of C peptide positive cells（P0.05） and the increased mRNA expressions of pancreas development related transcription factor PDX-1,Ngn3,NeuroD1,MafA,insulin,Glut-2（P0.05）.Meanwhile,by the improved protocol the protein expressions of C peptide,PDX-1 and Nkx6.1 were positive,and insulin and C-peptide release were promoted（P0.05） as demonstrated by glucose challenge test.Conclusion INGAP-pp may facilitate the differentiation of hUCMSCs into IPCs.However,the IPCs are not as mature as normal human islet β cells.

关 键 词：脐带间充质干细胞 胰岛素分泌细胞 胰岛新生相关蛋白肽 分化 

分 类 号：R392.33[医药卫生—免疫学] R392.12[医药卫生—基础医学]