不同培养基对H9亚型禽流感病毒在MDCK细胞上的增殖效果研究Proliferation of Avian Influenza Virus H9 Subtype in MDCK Cells in Different Culture Media
张建伟;史爱华;沈佳;景小冬;章振华;李林;姜北宇;
摘要(Abstract):
[目的]比较3种培养基对H9亚型禽流感病毒在MDCK细胞上的增殖效果。[方法]采用DMEM+10%血清、低血清培养基(MEM-MD-611)、无血清培养基(SFE4Mega)3种培养基制备MDCK细胞单层,分别接种不同稀释度的H9亚型禽流感病毒。然后,分别加入含10μg/ml胰蛋白酶的3种培养基作为维持液,培养病毒,每隔24 h观察其细胞病变,并测定上清HA滴度。[结果]在培养72~96 h时,无血清培养基病毒液的HA滴度高于低血清培养基,而低血清培养基则高于含血清培养基。[结论]3种培养基均可用于制备禽流感病毒抗原,其中无血清培养基、低血清培养基更适用于流感病毒抗原的制备。
关键词(KeyWords): 培养基;H9亚型流感病毒;MDCK细胞;增殖效果
基金项目(Foundation): 北京市农林科学院一般项目(2010A007)
作者(Author): 张建伟;史爱华;沈佳;景小冬;章振华;李林;姜北宇;
Email:
DOI: 10.13989/j.cnki.0517-6611.2012.08.010
参考文献(References):
- [1]连京华李惠敏刘玉山.目前禽流感的流行特点及防控对策[J].家禽科学,2010(11):25-27.
- [2]杨涛,刘明,张云,等.重组H5N3禽流感病毒株在MDCK中大规模增殖条件研究[J].中国预防兽医学报,2007,29(3):199-203.
- [3]KISTNERO,BARRETT N,MUNDT W,et al.Development of a novel influ-enza vaccine derived from a continuous cell line[J].Vaccine,2001,18(1):50-54.
- [4]KISTNERO,BARRETT PN,MUNDT W,el al.Development of a mammali-an cell(Vero)derived candidate influenza virus vaccine[J].Vaccine,1998,16(9/10):960-968.
- [5]GENZEL Y M,FISCHER U.REICHL.Serum-free influenza virus produc-tion avoiding washing steps and medium exchange in large-scale micro-carrier culture[J].Vaccine,2006,24:3261-3272.
- [6]GENZEL Y,OLMER R M,SCHAFER B,et al.Wave microcarrier cultiva-tion of MDCK cells for influenza virus production in serum containing andserum-free media[J].Vaccine,2006,24:6074-6087.
- [7]MERTEN OW.Development of serum-free media for cell growth and pro-duction of viruses/viral vaccines-safety issues of animal products used inserum-free media[J].Dev Biol(Basel)2002,111:233-57.