MDCK单细胞悬浮生长的驯化筛选及其在AIV增殖上的初步应用  

Adaptation and screening of MDCK cells in single-cell suspension growth mode for AIV replication

作　　者：冯磊[1] 吴培培[1] 褚轩[1] 王伟峰[1] 陈丽[1] 侯继波[1] 

机构地区：[1]江苏省农业科学院国家兽用生物制品工程技术研究中心,江苏南京210014

出　　处：《浙江农业学报》2015年第6期913-920,共8页Acta Agriculturae Zhejiangensis

基　　金：农业部公益性行业(农业)科研专项经费项目"动物疫苗制剂及下游工艺技术研究与示范"(201303046);江苏省农业科技自主创新资金项目[SCX(13)5036]

摘　　要：采用逐步降低培养基中血清含量的方法获得可悬浮生长的MDCK-Sus细胞株,对其生长代谢情况、细胞膜表面病毒受体丰度,以及在生物反应器中悬浮培养该细胞对禽流感H9亚型病毒增殖进行了初步应用研究。结果表明,MDCK-Sus细胞株可完全适应无血清营养条件并呈单细胞悬浮生长状态,细胞生长旺盛,平均比生长速率达到0.556 d-1,最大活细胞密度达到2.42×106cells·m L-1,并可检测出其细胞膜表面具有正常丰度的唾液酸-α-2,3-半乳糖糖链受体。在MOI为0.05,TPCK处理的胰蛋白酶作用浓度为1μg·m L-1的条件下,AIV-H9亚型病毒JS03株可在MDCK-Sus细胞中获得较好的增殖HA效价,达到8log2·25μL-1。MDCK-Sus cell strain was adapted and screened out by reducing serum concentration in culture medium,the growth and metabolism status,abundance of avian influenza virus（ AIV） receptor on cell membrane were detected and AIV replication in bioreactor scale was primarily applied. The results showed that the average specific growth rate of 0. 556 d- 1and maximum viable cell density of 2. 42 × 106cells·m L- 1could be achieved in MDCK-Sus singlecell suspension growth mode in serum free media after adaptation and screening. Normal abundance of AIV receptor,sialic acid-α-2,3-gal,could be detected on MDCK-Sus cells membrane. AIV-H9 JS03 virus strain could be replicated with 8log2·25μL- 1of HA titer in MDCK-Sus cells suspension cultured in bioreactor with the condition of MOI0. 05 and 1μg·m L- 1TPCK treated proteinase.

关 键 词：MDCK细胞 禽流感病毒 单细胞悬浮培养 禽流感病毒受体 

分 类 号：S855.3[农业科学—临床兽医学] Q291[农业科学—兽医学]