Conophylline与尼克酰胺联合诱导脂肪源干细胞为功能性类胰岛细胞  

Differentiation of adipose-derived stem cells into functional islet-like cells using Conophlline and Nicotinamide

作　　者：杨萍[1] 郑勤龙 王劲峰[1] 黄家学[1,3,4] 

机构地区：[1]天津大学药物科学与技术学院,天津市300072 [2]北欧生物科技(北京)有限公司,北京市102206 [3]协和干细胞基因工程有限公司,天津市300384 [4]中源协和干细胞生物工程股份公司,天津市300050

出　　处：《中国组织工程研究》2013年第6期1024-1028,共5页Chinese Journal of Tissue Engineering Research

摘　　要：背景:1型糖尿病的胰岛移植治疗一直面临供体来源不足与免疫排斥两大关键问题,寻找一种自体来源的种子细胞通过组织工程方法制备类胰岛组织可以提供充足新型供体、降低异基因供体移植的不良反应。目的:分析成人脂肪干细胞体外分化为对葡萄糖敏感、可分泌胰岛素的功能性胰岛样细胞团的能力,探索体外制备类胰岛组织的技术路线。方法:首先分离纯化人体脂肪干细胞,采用新型植物诱导剂Conophylline与其他诱导因子的不同组合将脂肪干细胞诱导分化为胰岛样细胞团,观察不同组合的诱导分化效率,并利用特异性染色、RT-PCR,免疫细胞化学等方法对诱导分化后的细胞团在基因水平与蛋白水平上进行鉴定,最后用ELISA法检测细胞团在不同浓度葡萄糖刺激下胰岛素的分泌情况。结果与结论:脂肪干细胞具有多能干细胞特性,可诱导分化为具有胰岛素分泌和葡萄糖浓度反应性类胰岛细胞团;Conophylline与尼克酰胺联合诱导可大幅度提高诱导分化效率。BACKGROUND： AIIogenic islet transplantation for type 1 diabetes is limited by shortage of donor islet cells and immune suppression, bioengineered islet-like clusters developed from autogenic stem cells might benefit diabetic patients by immunorejection free and off-shelf cellular treatment. OBJECTIVE： To investigate whether adipose tissue from adults have the potential to differentiate into insulin secreting, glucose responsive, functional islet-like clusters and to develop a practical approach for islet bioengineering in vitro. METHODS： Human adipose stem cells were isolated, cultured and expanded from human subcutaneous adipose tissue. Conophylline, a new plant derived inducer, in combination with other factors were used to induce human adipose stem cells into islet-like clusters and their efficiencies for islet-like clusters generation were compared. Differentiated cells were identified by tissue specific surface marker staining, reverse transcription-PCR and immunocytochemistry at protein and mRNA level. Finally, insulin secretion of cell clusters in different glucose concentrations was tested with enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION： Multipotential adipose stem ceils were cultured and expanded from adult subcutaneous adipose tissues. Those stem cells were conveniently differentiated into glucose responsive and insulin-secreting functional islet-like clusters. Conophylline in combination with Nicotinamide have the best efficiency of differentiating human adipose stern cells into islet cells.

关 键 词：干细胞 脂肪干细胞 尼克酰胺 Conophylline 胰岛样细胞团 糖尿病 胰岛素 分化 高效诱导 胰岛分化 干细胞图片文章 

分 类 号：R394.2[医药卫生—医学遗传学]