壳寡糖作为基因载体的体外评价

In Vitro Evaluation of Chito-oligosaccharides as Gene Vectors

目的评价壳寡糖作为基因载体的理化性质及转染293T细胞的活性。方法选用相对分子质量小的壳寡糖,使用噻唑蓝(MTT)法检测不同浓度壳寡糖的细胞毒性,琼脂糖凝胶电泳检测壳寡糖结合及保护质粒DNA的作用,荧光显微镜观测转染结果。结果高浓度壳寡糖第2天时具有较低毒性,第4天时细胞存活率高于不做处理的对照细胞;质量比>20倍时,壳寡糖几乎完全结合DNA,并且具有一定保护的作用;转染293T细胞后,荧光显微镜能够观测到表达的绿色荧光蛋白。结论壳寡糖可以作为基因载体转染293T细胞。

Objective To evaluate the physicochemical property of chito-oligosaceharides and the activity of the transfected 293T cells. Methods Chito-oligosaccharides of small relative molecular mass was used in our study. The cytotoxicity of different chito-oligosaccharides was measured by MTT. The capability of chito-oligosaccharides binding and protecting plasmid DNA was detected with agarose gel eleetrophoresis. The result of transfection was observed under fluorescence microscope. Results On the second day, high concentration of chito-oligosaccharides demonstrated little toxicity. On the fourth day, the cell survival rate of all experimental groups was higher than that of the control group. Chito-oligosaccharides exhibited complete binding with all DNA and effective protection when the mass ratio was over 20. Green fluorescence was found after transfection. Conclusion Chito-oligosaccharides was used as gene verors to transfect 293T cells.