| H3N2流感病毒冷适应株的无血清培养 |
| 投稿时间：2015-01-19 修订日期：2015-02-03 点此下载全文 |
| 引用本文：郭琦,耿兴良,龙云凤,杨帆,宋绍辉,刘婧,戴宗祥,姜述德,廖国阳.H3N2流感病毒冷适应株的无血清培养[J].医学研究杂志,2015,44(7):26-29 |
| DOI：
10.11969/j.issn.1673-548X.2015.07.008 |
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| 基金项目: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) |
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| 中文摘要: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H3N2流感病毒冷适应株无血清培养的适宜条件。 |
| 中文关键词:无血清培养 流感病毒冷适应株 Vero细胞 |
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| Cultivation of Cold-adapted Influenza Virus Strain H3N2 with Serum-free Medium |
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| Abstract: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 influenza virus strain H3N2 in serum-free media has been optimized. |
| keywords:Serum-free medium Cold-adapted influenza virus Vero cells |
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