人脐带间充质干细胞分化胰岛样细胞过程中胰岛素和巢蛋白的表达

发布时间：2017-12-26 18:07

本文关键词：人脐带间充质干细胞分化胰岛样细胞过程中胰岛素和巢蛋白的表达　出处：《中国组织工程研究》2016年50期 　论文类型：期刊论文

【摘要】：背景:国内外研究证实脐带间充质干细胞可定向诱导分化为胰岛样细胞,但分化过程中胰岛素与巢蛋白表达的变化等相关研究甚少。目的:观察人脐带间充质干细胞向胰岛素样细胞分化过程中胰岛素与巢蛋白表达的变化。方法:采用UltraCULTURE培养基在体外培养人脐带间充质干细胞,取培养的第3代细胞,按照两阶段诱导方案将其定向诱导分化为胰岛样细胞,第1阶段:在UltraCULTURE培养基中加入4 nmol/L活化素A,25μg/L表皮生长因子,100μg/Lβ-神经生长因子,10 mmol/L尼克酰胺,诱导培养至14 d。第2阶段:在UltraCULTURE培养基中加入1%胰岛素-转铁蛋白-硒,10 mmol/L尼克酰胺,10μg/L碱性成纤维细胞生长因子,继续诱导培养14 d。在诱导分化过程中,采用流式细胞技术检测分化细胞巢蛋白及胰岛素的表达,双硫腙染色鉴定胰岛样细胞团中锌离子的表达。结果与结论:(1)诱导分化为胰岛样细胞过程中胰岛素水平逐渐增高,至28 d达到较高水平,未诱导组胰岛素表达呈阴性;(2)诱导分化第14天,巢蛋白表达水平达到最大,随着诱导时间延长表达水平逐渐下降;(3)脐带间充质干细胞诱导28 d形成胰岛样细胞团,双硫腙染色阳性;(4)实验将脐带间充质干细胞成功诱导分化为胰岛样细胞,并且随着定向分化细胞的变化,细胞胰岛素与巢蛋白表达水平也出现相应的变化。
[Abstract]:Background: domestic and foreign studies confirm that umbilical cord mesenchymal stem cells can be induced to differentiate into islet like cells, but there is little research about the change of insulin and nestin expression during differentiation. Objective: To observe the changes in the expression of insulin and nestin in human umbilical cord mesenchymal stem cells (MSCs) to insulin-like cells. Methods: UltraCULTURE cultured human umbilical cord mesenchymal stem cells in vitro culture medium, cultured the cells of the third generation, according to the two phase of induced directionally induced BMSCs into islet like cells, the first stage: in 4 nmol/L activin A added UltraCULTURE medium, 25 g/L of epidermal growth factor, 100 g/L nerve growth factor beta, 10 mmol/L nicotinamide, cultured to 14 d. The second stage: after adding 1% insulin transferrin selenium UltraCULTURE medium, 10 mmol/L nicotinamide, 10 g/L basic fibroblast growth factor, continued to be cultured 14 d. During induction and differentiation, the expression of nestin and insulin in differentiated cells was detected by flow cytometry, and the expression of zinc ions in islet like cell clusters was identified by Dithizone staining. Results and conclusion: (1) differentiation into islet like cells in insulin levels increased to 28 d reached a high level, non induced expression of insulin group were negative; (2) fourteenth days of differentiation, the expression of nestin reached the maximum with time of induction was decreased; (3) umbilical cord mesenchymal stem cells induced by 28 d into islet like cell clusters. Dithizone staining; (4) the umbilical cord mesenchymal stem cells were successfully induced to differentiate into islet like cells, and with the change of cell differentiation, expression of nestin and insulin levels also appeared the corresponding changes.
【作者单位】：北京大学航天临床医学院血液内分泌科;中国中医科学院中药所药理室;
【基金】：航天中心医院科研基金(KY-201003),课题名称:人骨髓间充质干细胞向胰岛细胞定向分化的研究~~
【分类号】：R329.2
【正文快照】： 文章快速阅读:利用体外微环境诱导人脐带间充质干细胞定向分化为胰岛样细胞的过程人脐带间充质干细胞的体外分离、培养与鉴定细胞形态学观察、流式细胞仪检测表面表型、成骨成脂诱导分化步骤1诱导第1阶段脐带间充质干细胞向神经样细胞分化步骤2诱导第2阶段细胞形态学观察、流

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