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目的提高重组腺病毒在细胞中,特别是缺乏柯萨奇腺病毒受体的细胞中的转染效率。方法将阳离子材料聚乙烯亚胺PEI与重组腺病毒通过电荷作用形成复合物,用激光粒度仪及电位分析仪测定其粒径和电位;以表达β半乳糖苷酶的重组腺病毒作为模型病毒,研究其在富含(A549)或者缺乏(MDCK、LLC)柯萨奇腺病毒受体细胞中的转染效率和毒性,并用荧光标记复合物,观察其进入细胞的能力。结果与聚乙烯亚胺结合形成复合物可明显促进腺病毒进入细胞的能力,从而提高腺病毒在各种细胞中的转染效率,同时发现小分子的PEI-2 kD由于其较好的生物相容性,对腺病毒进入细胞后的传递影响较小,所以最终的基因传递效率要显著优于PEI-25 kD;PEI-2 kD在本复合物中表现出的细胞毒性要小于PEI-25 kD。结论PEI-2 kD可以用于病毒载体和非病毒载体结合的基因传递研究。
Objective To improve transfection efficiency of recombinant adenovirus in cells, especially in cells lacking Coxsackie adenovirus receptor. Methods Polyethyleneimine (PEI) and cationic adenovirus were used as the complex to form the complex through the charge. The particle size and potential were determined by laser particle size analyzer and potentiometric analyzer. The recombinant adenovirus expressing β-galactosidase was used as a model virus, The efficiency and toxicity of transfection in Coxsackie Adeno-recipient cells enriched for (A549) or lacking (MDCK, LLC) were studied and its ability to enter cells was visualized with a fluorescently labeled complex. Results The formation of complex with polyethylenimine significantly enhanced the ability of adenoviruses to enter cells and thus enhanced the transfection efficiency of adenovirus in various cells. At the same time, it was found that small molecule PEI-2 kD, due to its better biological phase The final gene delivery efficiency was significantly better than PEI-25 kD; PEI-2 kD exhibited cytotoxicity in this complex to be less than that of PEI-25 kD . Conclusions PEI-2 kD can be used in gene delivery studies that combine the viral and non-viral vectors.