HEK293T细胞扩增重组腺病毒研究

Amplifi cation of recombinant adenovirus in HEK293T cells

摘要: 目的探讨HEK293T细胞扩增重组腺病毒的可行性。方法重组腺病毒保存液稀释后感染HEK293T细胞,观察细胞病变效应,半数组织培养感染量测定扩增病毒的滴度,通过体外感染心肌细胞,成纤维细胞验证其感染活性。结果 HEK293T细胞可以扩增带有绿色荧光蛋白基因的重组腺病毒,病毒滴度为5×106PFU/mL,扩增后的腺病毒可以感染体外培养的心肌细胞和成纤维细胞。结论 HEK293T细胞可有效扩增带有绿色荧光蛋白基因的重组腺病毒。

Abstract: Objective To determine if recombinant adenovirus can be amplified in HEK293T cells. Methods After HEK293T cells were infected with a preservation solution of adenovirus,its effect on cytopathy was observed.Infectivity of amplified adenovirus was assayed with 50% of tissue-culture infective dose.Infectivity of adenovirus was confirmed with infected myocardial cells and fibroblasts in vitro. Results HEK293T cells promoted generation of recombinant adenovirus with green fluorescent protein.The titer of adenoviruses could reach 5×106PFU/mL.Adenoviral particles could infect myocardial cells and fibroblasts in vitro. Conclusion Recombinant adenovirus with green fluorescent protein can be amplified in HEK293T cells.