体外诱导人脐带间充质干细胞向胰岛β样细胞的分化  

Differentiation of mesenchymal stem cells derived from human umbilical cord into islet beta-like cells in vitro

作　　者：于洪宇[1] 马春宇[2] 郑晓明[3] 

机构地区：[1]辽宁医学院护理学院,辽宁省锦州市121001 [2]辽宁医学院研究生院,辽宁省锦州市121001 [3]辽宁医学院附属一院内分泌科,辽宁省锦州市121001

出　　处：《中国组织工程研究与临床康复》2010年第19期3461-3464,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:体外实验中人们发现,常规诱导骨髓间充质干细胞分化的胰岛β样细胞由于种种原因限制了其进一步的应用。关于人脐带间充质干细胞是否可以成功诱导的胰岛β样细胞尚未见系统报道。目的:进一步验证人脐带间充质干细胞向胰岛β样细胞分化的可行性。方法:采用胶原酶消化法分离人脐带细胞进行贴壁培养;传2代后,用高浓度葡萄糖(25mmol/L)培养液DMEM(含体积分数为10%胎牛血清)以及碱性成纤维细胞和尼克酰胺诱导脐带间充质干细胞向胰岛β样细胞分化。倒置显微镜下观察间充质干细胞诱导后的形态变化,用胰岛β细胞特异染色方法双硫腙染色鉴定诱导后细胞游离锌离子浓度;免疫细胞化学鉴定诱导后细胞内是否储存有胰岛素。结果与结论:第2代脐带间充质干细胞经过高糖诱导后,间充质干细胞形成细胞团;并且形成了双硫腙染色阳性的细胞团;免疫细胞化学表明诱导后细胞内的细胞胰岛素染色阳性。结果表明人脐带分离出的间充质干细胞在体外可以定向诱导分化为胰岛β样细胞,这种胰岛β样细胞具有表达、储存胰岛素的功能。BACKGROUND：In vitro experiment has shown that conventional induction-induced islet β-like cells differentiated from bone marrow mesenchymal stem cells which limited its further application.The possibility of mesenchymal stem cells（MSCs） derived from human umbilical cord differentiating into islet β-like cells in vitro has not yet been professionally reported.OBJECTIVE：To verify the feasibility of differentiation of MSCs from human umbilical cord into islet β-like cells.METHODS：Human umbilical cord cells were cultured by collagen enzyme digestion method.Following passage 2,MSCs from umbilical cord were induced by high-concentration glucose（25 mmol/L） DMEM（containing 10% fetal bovine serum） as well as basic fibroblast and niacinamide to differentiate into islet β-like cells.Morphological changes in MSCs were observed following induction under an inverted microscope.We stained the clusters with dithizone which is a zinc-chelating agent known to selectively stain pancreatic β-cells,because of their high zinc content.Insulin in treated cells was examined by immunocytochemistry.RESULTS AND CONCLUSION：Following high-glucose induction,the second passage of MSCs from umbilical cord formed cell mass,and DTZ-stained cell cluster formed.Immunocytochemistry results have confirmed that these cell clusters were positive for insulin.Above-described results have suggested that MSCs derived from human umbilical vein can be induced into islet β-cells in vitro.These islet β-cells can synthesize and store insulin.

关 键 词：人脐带 间充质干细胞 诱导 胰岛素 细胞培养 

分 类 号：R394.2[医药卫生—医学遗传学]