Apelin-13促进脐带华通胶间充质干细胞分化成血管网

发布时间：2018-02-21 21:38

本文关键词： 间充质干细胞 apelin- 血管生成三维培养脐带　出处：《天津医药》2015年06期 　论文类型：期刊论文

【摘要】：目的探索apelin-13对干细胞定向血管网分化的调控作用。方法采用组织块贴壁法从人脐带华通胶组织中分离间充质干细胞,流式细胞仪检测其免疫表型。取第3代人华通胶间充质干细胞,分别接种于4个培养瓶中,记为A1、A2、A3、A4组,用诱导液诱导7 d,每天向4组中分别加入浓度0、1×10-6、10×10-6、100×10-6mol/L的apelin-13 20μL,倒置显微镜下观察细胞形态及生长情况,并对其进行血管性血友病因子(v WF)抗体免疫荧光染色及CD31流式细胞学鉴定。使用水凝胶的三维培养基对诱导后的内皮细胞进行培养,并按照原A1、A2、A3、A4组的顺序重新依次编号为S1、S2、S3、S4组。每天分别向S1、S2、S3、S4组中加入浓度0、1×10-6、10×10-6、100×10-6mol/L的apelin-13 20μL。7 d后倒置显微镜下观察细胞的形态、生长情况及在三维培养基中形成血管样结构情况。结果人华通胶间充质干细胞可以诱导分化为v WF免疫荧光染色阳性的内皮样细胞。随着apelin-13蛋白浓度的增加,CD31阳性表达率也随之升高。显微镜下观察到内皮样细胞在水凝胶的三维培养基中能够形成血管样结构。结论证实apelin-13参与并调控人脐带华通胶间充质干细胞分化形成血管网。
[Abstract]:Objective to investigate the effect of apelin-13 on the differentiation of stem cell directed vascular network. Methods Mesenchymal stem cells were isolated from human umbilical cord Huatong glue tissue by tissue mass adherence method. Flow cytometry was used to detect its immunophenotype. Mesenchymal stem cells (MSCs) of the third generation of human Huatong gum were inoculated in 4 culture flasks and recorded as A1A2A2A3A4 group. After induction with the inducer for 7 days, apelin-13 20 渭 L of 10 脳 10 -6 mol / L 10 脳 10 -6 渭 L apelin-13 was added to the 4 groups each day. The morphology and growth of the cells were observed under inverted microscope. The vascular Willebrand factor v WF antibody was detected by immunofluorescence staining and CD31 flow cytology. The induced endothelial cells were cultured in hydrogel culture medium. In the same order, the former A1A1A2OA3A4 group was renumbered as S1OS2S2OS3OS4 group. The morphology of the cells was observed under inverted microscope after 20 渭 7d after adding 10 脳 10-6U 10 脳 10-6U 10 脳 10-6mol / L apelin-13 20 渭 7d to the S1OS2S2OS3 S4 group, respectively, in the order of the original A1A1A2A2A3A4 group, and the order of the former A1A1A2A2A3A4 group was as follows. Results the mesenchymal stem cells of human Huatong gum could be induced to differentiate into vWF-positive endothelial cells. With the increase of apelin-13 protein concentration, CD31 was positive. It was observed under microscope that endothelioid cells could form vascular-like structure in hydrogel culture medium. Conclusion it is confirmed that apelin-13 participates in and regulates the differentiation of mesenchymal stem cells from human umbilical cord Huatong gum to form vascular network.
【作者单位】：安徽医科大学海军临床学院;海军总医院心脏中心;
【基金】：国家自然科学基金资助项目(81170094)
【分类号】：R329.2

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