肠道病毒71型在RD细胞和Vero细胞中的增殖动力学特征  

Proliferative Kinetics Characteristics of Enterovirus Type 71 in Rhabdomyosarcoma Cells and Vero Cells

作　　者：齐向云[1,2] 李康[2,3] 夏燕平[2,4] 杨静[2] 戴岳[1] 

机构地区：[1]中国药科大学中药药理教研室,江苏南京211198 [2]军事医学科学院放射与辐射医学研究所,北京100850 [3]北京工业大学生命科学与生物工程学院,北京100124 [4]河南中医学院药学院,河南郑州450008

出　　处：《生物技术通讯》2014年第3期333-336,共4页Letters in Biotechnology

基　　金：重大新药创制重大专项(2013ZX09304102);国家高技术研究发展计划(2012AA022501)

摘　　要：目的:利用噬斑法比较肠道病毒71型(EV71)在RD细胞和Vero细胞中的增殖动力学特征。方法:首先探讨培养基类型、羟乙基哌嗪乙磺酸(HEPES)、胎牛血清(FBS)、牛血清白蛋白(BSA)及甲基纤维素(MC)含量对EV71噬斑形成的影响,得到最适营养覆盖物配比;进一步,EV71以感染复数(MOI)为0.1分别接种RD细胞和Vero细胞,收集接种后不同时间点的细胞培养液,噬斑法测定各时间点培养液上清中的病毒滴度,并绘制log2(病毒滴度)-时间图,对比分析EV71在2种细胞中的增殖动力学特征。结果:终浓度含1%MC和2%FBS的MEM(1×)或DMEM(1×)为EV71噬斑形成的最适营养覆盖物;EV71在RD细胞和Vero细胞中的增殖周期均约为12 h,MOI=0.1时,EV71在RD细胞中的增殖活动较Vero细胞中活跃,增殖效率比Vero细胞中高2个数量级。结论:用RD细胞扩增EV71比Vero细胞更具优势。Objective： To study the proliferative kinetics characteristics of enterovirus type 71（EV71） in rhabdomyosarcoma（RD） cells and Veto cells, respectively. Methods： Firstly, the culture media type, the present of hydroxyethyl piperazine ethanesulfonic aeid（HEPES）, fetal bovine serum（FBS） and bovine serum alhumin（BSA）, and the concentration of methyl cellulose （MC） were respectively investigated to obtain the fit cultural slipcover type for the plaque assay of EV71. Then, RD cells and Vero cells were cultured with EV71 at a multiplicity of infection（MOI） of 0.1 for 1 h and then with EV71-free culture media（with 2% FBS） at 37℃, 5% CO2. The cell culture fluid was collected at a series of time points post infection and the proliferative kinetics characteristics of EV71 in RD cells and Veto cells were respectively determined by measuring the virus titers of the culture media supernatant at each time points through plaque assay. Results： Cultural slipcover with 1% MC, 2% FBS, MEM （1x） or DMEM（1x） was the certain slipcover. EV71 had a multiplication cycle of about 12 hours in both RD cells and Vero cells. And when infected with EV71 at an MOI=0.1, the final viral titer in culture media supernatant of RD cells was 100 times as of Vero cells. Conclusion： It＇s more predominant to use RD cells for EV71 amplification than Vero cells.

关 键 词：肠道病毒71型 噬斑 增殖动力学特征 RD细胞 VERO细胞 

分 类 号：R373.2[医药卫生—病原生物学]