人脐带间充质干细胞体外向胰岛样细胞诱导分化及其治疗糖尿病效果  

Differentiation of human umbilical cord blood mesenchymal stem cells into islet-like cells in vitro and its effects on diabetes

作　　者：李俊林[1] 李德华[1] 赵宝东[1] 王长辉[1] 王志云[1] 裴丹[1] 

机构地区：[1]辽宁医学院解剖学教研室,辽宁省锦州市100021

出　　处：《中国组织工程研究与临床康复》2009年第14期2636-2640,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:相对于其他来源的干细胞而言,脐带间充质干细胞免疫排斥反应和免疫原性较小,但并不等同于无任何免疫排斥反应。目的:观察人脐带间充质干细胞体外向胰岛样细胞的诱导分化,及其移植后对糖尿病大鼠的治疗效果。设计、时间及地点:细胞学体内对照观察,于2008-11在辽宁医学院解剖学教研室实验室完成。材料:清洁级雄性SD大鼠36只,随机分为4组:胰岛样细胞组12只、间充质干细胞组12只、模型对照组6只、正常对照组6只。人脐带间充质干细胞由天津国家干细胞中心提供。方法:取传至第3代的人脐带间充质干细胞,待细胞融合至70%～80%后,换用含碱性成纤维细胞生长因子、尼克酰胺的DMEM高糖培养基,向胰岛样细胞诱导分化。除正常对照组外,其余3组大鼠均腹腔注射链脲佐菌素建立糖尿病模型。造模后,胰岛样细胞组于肾被膜下植入胰岛样细胞2×106个,间充质干细胞组同法植入等量未诱导的脐带间充质干细胞,模型对照组植入不含任何细胞的培养液1mL。主要观察指标:诱导后胰岛样细胞的鉴定,胰岛样细胞的移植效果。结果:诱导前脐带间充质干细胞呈长梭形贴壁生长;诱导4d后细胞向中央聚集,出现胰岛样细胞团,此后胰岛样细胞团逐渐增大且数量增多;诱导7d双硫腙染色呈阳性表达。移植后2周,胰岛样细胞组血糖浓度明显降低,一直维持至第6周;间充质干细胞组血糖浓度在移植后很快下降,但4周后血糖浓度上升;模型对照组血糖浓度一直处于糖尿病血糖浓度范围内。免疫组化染色结果显示,移植后4周胰岛样细胞组胰腺管腔内可见大量胰岛素表达,间充质干细胞组仅有表达少量胰岛素,正常对照组胰岛素表达无明显变化。结论:人脐带间充质干细胞体外可诱导分化为胰岛样细胞。与脐带间充质干细胞相比,胰岛样细胞能够较长时间维持大鼠血糖浓度在正常水平,BACKGROUND： Compared with other stem cells, immunological rejection and immunogenicity of umbilical cord blood mesenchymal stem cells （UCB-MSCs） are small. OBJECTIVE： To observe in vitro differentiation of UCB-MSCs into islet cells and the therapeutic efficacy on diabetic rats following transplantation. DESIGN, TIME AND SETTING： The cytology in vivo controlled study was performed at the Laboratory of Department of Anatomy, Liaoning Medical University in November 2008. MATERIALS： A total of 36 clean male Sprague Dawley rats were randomly assigned into 4 groups, islet cell group （n=12）, MSC group （n=12）, model control group （n=6）, and normal control group （n=6）. UCB-MSCs were obtained from the National Stem Cell Center, Tianjin. METHODS： Human UCB-MSCs at passage 3 were obtained. When the cell fusion 70 to 80 percent, UCB-MSCs were incubated in high-glucose DMEM containing basic fibroblast growth factor and nicotinamide, and differentiated into islet-like cell clusters. Except the normal control group, rats in other groups were intraperitoneally injected with streptozotocin to establish diabetes models. After model induction, rats in the islet-like cell group were implanted with islet-like cells （2×10^6） under the renal capsule. Rats in the MSC group were treated with the same volume of non-induced UCB-MSCs. Rats in the model control group were subjected with 1 mL medium without any cells. MAIN OUTCOME MEASURES： Identification of islet-like cells after induction; transplantation efficiency of islet-like cells. RESULTS： Before induction, UCB-MSCs were spindle and adhered to the wall. At 4 days following induction, ceils centralized, and islet-like cell clusters appeared. Subsequently, islet-like cell clusters gradually increased. At 7 days following induction, dithizone staining were positive. At 2 weeks following transplantation, blood glucose concentration in the islet-like cell group significantly reduced, and maintained till 6 weeks. The blood glucose concentration in the M

关 键 词：脐带间充质干细胞 诱导 胰岛样细胞 糖尿病 

分 类 号：R394.2[医药卫生—医学遗传学]