中国组织工程研究与临床康复2011，Vol.15Issue(1)：87-91,5.DOI:10.3969/j.issn.1673-8225.2011.01.019
siRNA阻断B7H1在人胎盘间充质干细胞上表达对其黏附和迁移的影响
Effects of siRNA target gene B7H1 on adhesion and migration of human placenta mesenchymal stem cells
摘要
Abstract
BACKGROUND: Human placenta mesenchymal stem cells (hPMSCs) express costimulatory molecules B7H1 which plays an important role in its immunosuppressive capabilities. However, the effect of B7H1 on the biological behavior of hPMSCs has not been reported.OBJECTIVE: To investigate the effect of B7H1 blockade on adhesion and migration of hPMSCs.METHODS: hPMSCs were isolated from mature placenta by the method of digestion. Then in vitro hPMSCs were cultured,expanded and were used in test after third passage. Flow cytometry and RT-PCR analysis were used to investigate the expression of B7H1 on hPMSCs. Specific siRNAs of B7H1 were transfected into hPMSCs via cathodolyte liposome transfection method. RT-PCR was used to detect the changes of B7H1 gene expression. Then the adhesion of hPMSCs was determined by cell count, and the migration of hPMSCs was evaluated using Transwell method.RESULTS AND CONCLUSION: siRNA could effectively block the expression of B7H1 which highly expressed on hPMSCs. After seeded for half an hour, the cell adhesion rate had no significantly difference in the blockade group and the control group. For 1 hour or 3 hours, however, the adhesion rate of hPMSCs was significantly higher in the blockade group than in the control group (P ＜ 0.05). Transwell assay indicated that the migration numbers of hPMSCs in the blockade group were significantly less compared with the control group under the culture conditions of SDF-1α, DMEM-LG complete medium or hPMSCs culture supernatant (P ＜ 0.01). The results showed that the adhesion ability of hPMSCs was enhanced while the ability of migration was inhibited after B7H1 blockade and suggested that B7H1 plays an important role in the adhesion and migration of hPMSCs.关键词
人胎盘间充质干细胞/B7H1/siRNA/黏附/迁移分类
医药卫生引用本文复制引用
张思英,王国艳,都海波,栾希英..siRNA阻断B7H1在人胎盘间充质干细胞上表达对其黏附和迁移的影响[J].中国组织工程研究与临床康复,2011,15(1):87-91,5.基金项目
山东省自然科学基金资助项目(Y2006C02) （Y2006C02）
山东省医药卫生科技发展计划(2007HZ039) （2007HZ039）
滨州医学院科研启动基金(Y2007KYQD09). （Y2007KYQD09）
