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机构地区:[1]福建医科大学解剖学与组织胚胎学系,福州350004
出 处:《解剖学杂志》2008年第5期640-642,共3页Chinese Journal of Anatomy
基 金:教育部科学技术研究重点项目(01058);福建省科技厅重点项目(2002Y014);福州市科技局项目(2002-14)
摘 要:目的:研究新型阳离子聚合物Sofast基因转染试剂(Sofast)对大鼠骨髓间充质干细胞(BMSCs)进行基因修饰的可行性。方法:在体外分离和扩增MSCs,在Sofast介导下行pEGFP-N1转染BMSCs,倒置荧光显微镜下观察转染效率及瞬时表达情况,MTT试验检测转染细胞的存活率。结果:增强性绿色荧光蛋白(EGFP)在基因转染24 h后开始表达,48~72 h最高。1周后表达逐渐减弱;质粒与Sofast体积比例为3∶1时,pEGFP-N1转染BMSCs的效率为85%,细胞存活率为94.2%。结论:Sofast介导EGFP转染BMSCs是一种较好的转染标记方法。Objective: To study the feasibility of cationic polymer Sofast transfection reagent into bone marrow mesenchymal stem cells (BMSCs). Methods: Enhanced green fluorescent protein (EGFP) was transfected with Sofast into BMSCs, isolated and amplified in vitro. Transfection efficiency and transient expression were evaluated by inverted fluorescent microscope. The vitality of transfected cells was detected by MTT test. Results: The expression of EGFP began in 24 hours after transferring, reached maximum in 48-72 hours and decreased after one week. When the volume ratio of plasmid and Sofast was 3 : 1, the efficiency of transferring pEGFP-N1 into BMSCs could achieve to 85%. The cell vitality was 94.2%. Conclusion: Sofast, used to mediate EGFP transfecting BMSCs, is a better label method.
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