EGFP基因转染骨髓间充质干细胞的实验研究.doc

EGFP基因转染骨髓间充质干细胞的实验研究
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作者:郑立恒魏会平许松梅何波杨明理王黎明刘全胜【摘要】目的: 探讨外源性EGFP基因转染修饰兔骨髓间充质干细胞(m esenchymal stem cells, MSCs)的可行性。方法: 用增强型荧光绿色蛋白(enhanced green fluorescent protein, EGFP)作为报告基因,以脂质体法转染骨髓间充质干细胞,观察转染结果、表达情况及对靶细胞活力的影响。结果: 经荧光显微镜下观察证实:成功地对骨髓间充质干细胞实现了基因转染。结论: 兔骨髓间充质干细胞能够在体外适宜的条件下进行长期培养; 骨髓间充质干细胞可直接作为基因靶细胞, 建立稳定表达EGFP 的骨髓间充质干细胞系具有可行性,为进一步做标记的MSCs细胞的移植研究奠定基础。
【关键词】增强型绿色荧光蛋白(EGFP);转染;骨髓间充质干细胞(MSCs)
【ABSTRACT】 Objective: To study the feasibility of exogenous gene transfecting into bone marrow mesenchymal stem cells (MSCs). Methods: MSCs were cultured and treated with pEGFPC1 with the help of lipofectamine. The transgene results were confirmed by fluoroscopy and the cell viability after transfection was evaluated by light microscopy and electron microscopy. Results: Bone marrow MSCs were essfully transfected with pEGFPC1 gene. Conclusion: The MSCs originated from rat bone can be cultured in vitro under appropriate conditions, and they can be directly used as gene target cells. Moreover, it is feasible to construct the MSCs expressing EGFP stably, which can be further applied to label target cells in the transplantation study.
【KEY WORDS】 Enhanced green fluorescent protein(EGFP); Transfection; Mesenchymal stem cells(MSCs)
间充质干细胞(mesenchymal stem cells, MSCs)由于具有自我更新,多向分化的潜能,且容易获取和体外扩增培养,成为近年来干细胞研究的热点。迄今为止,国内外已对MSCs培养、生物学特性及治疗应用等进行了大