人脐带间充质干细胞分化为胰岛β细胞的实验研究  

Experimental Research on Human Umbilical Cord Wharton′s Jelly-Derived Mesenchymal Stem Cells Differentiate into Insulin-Producing Cells in Vitro

作　　者：马桂霞[1] 何红燕[2] 罗敏洁[3] 黄天华[3] 肖升平[1] 崔冰琳[1] 马廉[1] 

机构地区：[1]汕头大学医学院第二附属医院儿科,广东汕头515041 [2]汕头大学医学院第一附属医院外科,广东汕头515031 [3]汕头大学医学院生殖中心实验室,广东汕头515031

出　　处：《实用儿科临床杂志》2008年第8期618-620,共3页Journal of Applied Clinical Pediatrics

基　　金：广东省社会发展计划项目资助(2006B3600502)

摘　　要：目的探讨人脐带华尔通胶质来源的间充质干细胞(MSCs)分化为胰岛素分泌细胞的可能性。方法从足月剖宫产的新生儿脐带华尔通胶质中分离培养出MSCs,传至第5代用含尼克酰胺和β巯基乙醇的低糖达尔伯克必需基本培养基预诱导,以含尼克酰胺和β巯基乙醇的高糖达尔伯克必需培养基诱导其向胰岛素分泌细胞分化,应用免疫细胞化学法检测其胰岛素的表达,以双硫腙染色鉴定胰岛β细胞,并用放射免疫法检测诱导后细胞培养液中胰岛素水平,检测均以未诱导细胞作为实验对照。结果免疫细胞化学法检测结果显示,经尼克酰胺和β巯基乙醇联合诱导后的人脐带MSCs表达胰岛β细胞的标记胰岛素。双硫腙染色发现脐带华尔通胶质来源的MSCs经尼克酰胺和β巯基乙醇诱导后细胞内锌离子增加。通过胰岛素放射免疫法检测诱导前后细胞培养液中胰岛素水平,实验组和对照组分别为(6.63±1.80)U/L和(3.39±0.21)U/L,差异具有统计学意义(P<0.05)。结论人脐带MSCs经尼克酰胺和β巯基乙醇联合诱导后可表达胰岛素β细胞的表面标记,增加细胞内锌离子水平,使之具备胰岛β细胞的特点,诱导后的细胞也可分泌胰岛素。用尼克酰胺和β巯基乙醇可诱导人脐带MSCs分化成为胰岛素分泌细胞,从而为1型糖尿病的细胞移植治疗提供新的细胞来源。Objective To explore the possibility of inducing human umbilical cord Wharton＇s Jelly - derived mesenchymal stern cells（MSCs） by nicotimine and β - mercaptoethanol to differentiate into insulin - producing cells. Methods MSCs were cultured from the Wharton＇s Jelly taken from full - terra section birth. MSCs of generation 5 were preinduced by low carbohydrates dulbecco＇s minimum essential medium within ni-cotimide and β - mercaptoethanol, then induced by high glucose dulbecco＇s minimum essential medium within nicotimine and β - mercaptoethanol. The expression of insulin marker was shown by immunocytochemistry, Zincum in cells was dyed by dithizone, and the levels of insulin in the media were detected by radioimmunoasay. Uncultured MSCs cells were as control group. Results The result of immunocytochemistry showed that MSCs induced by nicotimine and β -mercaptoethanol could express insulin,the marker of β - cells were in pancreatic islet. Zincum in those ceils derived from human umbilical cord Wharton＇s Jelly dyed by dithizone had an increase when they were induced by nicotimine and β - mercaptoethanol. Radioimmunoasay of insulin showed that the levels of insulin in experimental group and control group were （6.63 ± 1.80） U/L and （ 3.39 ± 0. 21 ） U/L, and the difference had statistical significance（P 〈0.05 ）. Conclusions MSCs derived from human umbilical cord Whwrton＇s Jelly expressed the marker of β cell of pancreatic islet after induced by nieotimine and β - mercaptoethanol. And the level of zineum in the differentiated cells were more increased than that in the undifferentiated cells. Much more,insulin was detected in the media of the defferentiated cells. This study showed that nicotimine and β - mercaptoethanol could induce the MSCs into insulin producing cells, thereby a new rescource of cell transplantion could be provided to cure type 1 diabetes mellitus.

关 键 词：脐带 间充质干细胞 胰岛 尼克酰胺 

分 类 号：R587.1[医药卫生—内分泌]