H3N2流感病毒冷适应株的无血清培养  

Cultivation of Cold-adapted Influenza Virus Strain H3N2 with Serum-free Medium

作　　者：郭琦[1] 耿兴良 龙云凤[1] 杨帆[1] 宋绍辉[1] 刘婧[1] 戴宗祥[1] 姜述德[1] 廖国阳[1] 

机构地区：[1]中国医学科学院/北京协和医学院医学生物学研究所,昆明650118

出　　处：《医学研究杂志》2015年第7期26-29,共4页Journal of Medical Research

基　　金：国家高技术研究发展计划("863"计划)项目(2012AA02A404);国家科技部国际科技合作项目(2011DFR30420)

摘　　要：目的：探索无血清培养H3N2流感病毒冷适应株的适宜条件。方法以无血清适应的Vero细胞为介质，对流感病毒冷适应株（H3N2）的培养条件进行优化，以得到较优的基础培养基、pH值、TPCK胰酶添加量及其补加时间、病毒接种量和病毒收获时间。结果 H3N2流感病毒冷适应株能够感染无血清适应的Vero细胞；经过优化，病毒培养液适宜pH为7．15-7．64， BSA和TPCK胰酶添加量均为0．8-1．2μg／ml，细胞培养36h接种病毒，接种量MOI为0．05-0．10，于25℃培养168h收获时病毒效价达最高。结论优化了H3 N2流感病毒冷适应株无血清培养的适宜条件。Objective To explore a better serum-free culture condition for cold-adapted influenza virus strain H3N2.Methods By using serum-free adapted Vero cells as medium, we optimized the culture condition of cold-adapted influenza virus H3N2 including appropriate basal medium, pH values, supplementary volume and time of TPCK-treated trypsin, inoculum and harvest time of virus. Results The cold-adapted influenza virus strain H3N2 can infect the serum-free cultured Vero cells.Through the experiments we found the optimum pH value of virus culture solution was 7.15-7.64 and supplementary final concentration of BSA and TPCK-treated trypsin was 0.8-1.2μg/ml respectively.If the virus inoculate in Vero cells which have been cultivated for 36h with 0.05-0.10 MOI and been cultured for 168h at 25℃, the virus will achieve the highest titer.Conclusion The culture conditions of the cold-adapted in-fluenza virus strain H3N2 in serum-free media has been optimized.

关 键 词：无血清培养 流感病毒冷适应株 VERO细胞 

分 类 号：R3[医药卫生—基础医学]