miR-21促进脐带血间充质干细胞血管向分化的体外研究

doi:10.19439/j.sjos.2017.05.002

上海口腔医学 ›› 2017, Vol. 26 ›› Issue (5): 471-475.doi: 10.19439/j.sjos.2017.05.002

miR-21促进脐带血间充质干细胞血管向分化的体外研究

田忠奎, 刘琳, 陈冬雷, 王培, 徐景明, 汤易, 汤春波

南京医科大学口腔医学研究所,南京医科大学附属口腔医院种植修复科,江苏南京 210029

收稿日期:2017-01-09 修回日期:2017-05-26 出版日期:2017-10-25 发布日期:2017-11-23
通讯作者: 汤春波, E-mail: cbtang@njmu.edu.cn
作者简介:田忠奎（1978-）,男,主治医师,E-mail:42341534@qq.com
基金资助:
国家自然科学基金 (81470778）; 江苏高校优势学科建设工程资助项目(2014-37)

Effect of miR-21 in promoting angiogenesis differentiation of umbilical cord blood mesenchymal stem cells in vitro

TIAN Zhong-kui, LIU Lin, CHEN Dong-lei, WANG Pei, XU Jing-ming, TANG Yi, TANG Chun-bo

Jiangsu Key Laboratory of Oral Diseases; Department of Implantology, Affiliated Hospital of Stomatology, Nanjing Medical University.Nanjing 210029, Jiangsu Province, China

Received:2017-01-09 Revised:2017-05-26 Online:2017-10-25 Published:2017-11-23

摘要/Abstract

摘要： 目的体外研究miR-21对脐带血间充质干细胞（umbilical cord blood derived mesenchymal stem cells,UCBMSCs）血管向分化的促进作用。方法培养UCBMSCs （来源于同济大学干细胞库）;构建Lenti-LacZ-Luciferase和Lenti-miR-21-Luciferase慢病毒载体;目的基因转染UCBMSCs后0、1、4、7及14d,分别采用实时荧光定量PCR及Western 印迹法检测关键成血管因子的表达。用第3代UCBMSCs分别转染Lenti-LacZ-Luciferase和Lenti-miR-21-Luciferase后,接种于Matrigel上培养,显微镜下观察其管样结构形成情况。采用SPSS13.0软件包对数据进行统计学分析。结果成功培养脐带血骨髓间充质干细胞并成功转染目的基因; qPCR以及Western 印迹法结果表明,miR-21组HIF-1α和VEGF在第4天开始显著高表达,并持续到7 d和14 d;Matrigel结果显示,Lenti-miR-21-Luciferase组管状结构最多,长度最长。结论MiR-21具有促进UCBMSCs血管向分化的作用。

关键词: miR-21, UCBMSCs, 血管再生

Abstract: PURPOSE: To investigate the effect of miR-21 on promoting vascular differentiation of umbilical cord blood derived mesenchymal stem cells (UCBMSCs) in vitro. METHODS: UCBMSCs (from Stem Cell Bank of Tongji University) were cultured; Lenti-LacZ-Luciferase and Lenti-miR-21-Luciferase Lentiviral vector were reconstructed; Real-time fluorescence quantification PCR and Western blot were performed to detect the expression of key angiogenic factors at 0, 1, 4, 7 and 14 d after target gene was transduced into UCBMSCs; UCBMSCs of the third generation were transduced with Lenti-LacZ-Luciferase and Lenti-miR-21- Luciferase; matrigel tube assay was performed to observe the tube-like structure under microscope. The data were statistically analyzed using Student's t test and variance analysis with SPSS13.0 software package. RESULTS: UCBMSCs were successfully cultured; The results of qPCR and Western blot showed the expression of HIF-1α and VEGF in miR-21 group significantly increased at 4 d and continued until 7 d and 14 d; Matrigel results showed the tubular structures in Lenti-miR-21-Luciferase group were more than other groups both in amount and in length. CONCLUSIONS: MiR-21 can promote vascular differentiation of UCBMSCs.

Key words: miR-21, UCBMSCs, Angiogenesis

中图分类号:

R329

田忠奎, 刘琳, 陈冬雷, 王培, 徐景明, 汤易, 汤春波. miR-21促进脐带血间充质干细胞血管向分化的体外研究[J]. 上海口腔医学, 2017, 26(5): 471-475.

TIAN Zhong-kui, LIU Lin, CHEN Dong-lei, WANG Pei, XU Jing-ming, TANG Yi, TANG Chun-bo. Effect of miR-21 in promoting angiogenesis differentiation of umbilical cord blood mesenchymal stem cells in vitro[J]. Shanghai Journal of Stomatology, 2017, 26(5): 471-475.

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miR-21促进脐带血间充质干细胞血管向分化的体外研究

Effect of miR-21 in promoting angiogenesis differentiation of umbilical cord blood mesenchymal stem cells in vitro

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