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评价N,N,N-三甲基壳聚糖盐酸盐(TMC)及其纳米粒子对质粒DNA(pDNA)的负载及保护能力,考察了其纳米复合物对人类肝癌细胞株(HepG 2细胞)的转染能力.通过复凝聚法制备TMC/pDNA纳米粒子,并采用透射电镜及原子力显微镜表征粒子形态和粒径;采用凝胶阻滞分析观察其对pDNA的保护情况;采用四噻氮唑盐(MTT)比色法测定细胞毒性;以Lipofectamine 2000转染试剂作为阳性对照,检测其对HepG 2细胞的转染活性;采用荧光倒置显微镜观察转染情况.结果表明负载pDNA的TMC纳米粒子多呈球形,粒径为100~300 nm,能有效地包裹和保护基因不被DNaseⅠ酶消化;当TMC与pDNA的质量比为10∶1时,在48 h达到最高的转染效率.

参考文献

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