课题背景：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）和福州市科技局项目（2002-14）资助的“骨髓间质干细胞移植治疗脊髓损伤的实验研究”的子课题，前期实验表明骨髓间充质干细胞移植治疗脊髓损伤能明显改善脊髓的神经功能，移植的骨髓间充质干细胞能向损伤部位移行，并能分化为神经样细胞和分泌神经营养物质。

同行评价：目前骨髓间充质干细胞的基因转染技术问题还面临着一定的难关。病毒介导的基因转染技术转染率高，但具有潜在的生物风险和免疫原性；非病毒式，包括显微注射法、颗粒轰击法、电穿孔法、裸露DNA直接注射法、脂质体包埋放等载体虽然相对安全，但转染率低，已转入细胞内的DNA易被细胞内的DNA酶融解，导致目的基因不能在细胞内稳定存在和表达，又限制了非病毒介导法的应用。

应用要点：实验采用梭华-Sofast转染骨髓间充质干细胞。实验结果表明：①Sofast介导绿色荧光蛋白转染骨髓间充质干细胞的转染率较Jet PEI、Superfect 和Lipofectamine 2000等转染试剂的转染率高。②Sofast介导绿色荧光蛋白转染骨髓间充质干细胞后，骨髓间充质干细胞的存活分数较Jet PEI、Superfect 和Lipofectamine 2000等转染试剂的存活分数高。

摘要
目的：梭华-Sofast是新一代阳离子聚合物转染试剂，具有高效率转染所必备特征，如浓缩DNA，将DNA运送到细胞内，并使其在细胞核内释放等，且细胞毒性很低，很稳定，不被血清清除等特点。观察梭华-Sofast介导绿色荧光蛋白转染骨髓间充质干细胞的转染效果及细胞毒性。
方法：实验于2006-03-01/12-31在福建医科大学神经生物学研究中心完成。①实验材料：实验动物选用清洁级健康雄性SD大鼠10只，二三月龄，体质量150~250 g,由福建医科大学实验动物中心提供。实验过程中对动物处置符合动物伦理学标准。②实验方法：SD大鼠10只，用密度梯度离心及贴壁法相结合的方法分离纯化大鼠骨髓间充质干细胞，分别以Sofast、阳离子聚合物Jet PEI、Superfect和阳离子脂质体Lipofectamine 2000为转染试剂，转染绿色荧光蛋白入骨髓间充质干细胞，倒置荧光显微镜下检测转染细胞的绿色荧光蛋白基因表达，并通过MTT法检测转染细胞毒性。
结果：梭华-Sofast、阳离子聚合物Jet PEI、Superfect和阳离子脂质体Lipofectamine 2000介导的绿色荧光蛋白能有效转染入骨髓间充质干细胞，转染效率分别为（85.62±8.76 ）%、（73.34±7.32）%、（75.61±7.79）%和（76.62±8.21）%，梭 华-Sofast转染率最高（P < 0.05）；转染细胞的存活分数分别为(92.18±9.27)%、(86.98±8.54)%,(87.84±8.59)%和(87.17±8.61)%,梭华-Sofast细胞毒性最低（P < 0.05）。
结论：梭华-Sofast介导绿色荧光蛋白转染骨髓间充质干细胞具有高转染效率和低细胞毒性的优点。
关键词：Sofast;绿色荧光蛋白；骨髓间充质干细胞；转染

吴碧莲，陈少强，贾小力.梭华-Sofast介导绿色荧光蛋白转染骨髓间充质干细胞的转染效果[J].中国组织工程研究与临床康复，2008，12(16):3029-3032 [www.zglckf.com/zglckf/ejournal/upfiles/08-16/16k-3029(ps).pdf]

Transfection efficiency of Sofast by transfecting green fluorescent protein into bone marrow mesenchymal stem cells

Abstract

AIM：Sofast is a new kind of cationic polymer transfection reagent, characterized by high transfection efficency, such as condensing DNA. DNA is transported into cells, and releases in cellular nucleus, with low cytotoxicity, stability, cannot be cleaned with serum. This article investigates the transfection efficiency and cytotoxicity of green fluorescent protein (GFP) transfected bone marrow mesenchymal stem cells (BMSCs) by Sofast.
METHODS: Experiments were performed at the Research Center of Neurobiology of Fujian Medical University from March to December 2006. ①Ten clean healthy male SD rats aged two or three months (150-250 g) were provided by Experimental Animal Center of Fujian Medical University. Animal intervention met the Animal Ethical Standards. ②BMSCs obtained from SD rats were isolated and purified in vitro by Percoll density gradient centrifugation combined with adherent method. GFP was transfected into MSCs by Sofast, cationic polymer Jet PEI and Superfect and cationic liposome Lipofectamine 2000. Expression of GFP was examined by inverted fluorescence microscope and the cytotoxicity of the trasfected cells was determined with MTT assay.
RESULTS: The transfection efficiency of Sofast, cationic polymer Jet PEI, Superfect and cationic liposome Lipofectamine 2000 was about (85.62±8.76)%，(73.34±7.32)%，(75.61±7.79)% and (76.62±8.21)%，and the transfection efficiency of Sofast was the highest (P < 0.05); the exist score of the trasfected cells was about (92.18±9.27)%，（86.98±8.54）%,（87.84±8.59）% and （87.17±8.61）%, and the cytotoxicity of Sofast was the lowest （P < 0.05）.
CONCLUSION: Sofast is a high efficiency but low cell toxicity transfection by transfecting GFP into BMSCs.

Wu BL, Chen SQ, Jia XL.Transfection efficiency of Sofast by transfecting green fluorescent protein into bone marrow mesenchymal stem cells.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(16):3029-3032(China)
[www.zglckf.com/zglckf/ejournal/upfiles/08-16/16k-3029(ps).pdf]