MDCK细胞悬浮驯化及对H9亚型禽流感病毒敏感性的研究

为获得对H9亚型禽流感病毒敏感的纯悬浮MDCK细胞株,通过逐渐降低血清的方法对贴壁依赖的MDCK细胞进行了无血清纯悬浮驯化,最终获得了一株对H9亚型禽流感病毒敏感的纯悬浮细胞株MDCK-sus。通过测定表明,该细胞株能够迅速增殖,倍增时间为24h,使用H9亚型禽流感病毒BX13株感染该细胞,培养72h,培养液的病毒HA价可达到1∶512,每0.1mL病毒含量达到108.5EID50,为利用生物反应器大规模悬浮培养制备H9亚型禽流感病毒抗原奠定了基础。

Conversion of MDCK cell line to suspension culture and its sensitivity study to H9 subtype avian influenza virus

In order to obtain all suspension MDCK cell line sensitive to H9 subtype avian influenza virus, the adherent MDCK cells were domesticated in serum-free suspension by gradually decreasing serum levels and a pure suspension cell line MDCK-sus susceptible to H9 subtype avian influenza was finally obtained. The results indicate that the cell line can multiply rapidly with a doubling time of 24 hours. After 72 hours of incubation with H9 subtype avian influenza virus BX13 strain, the virus HA of the culture medium can reach 1:512, and the viral content can reach 108.5 EID50 every 0.1 mL, which laid a foundation for the production of H9 subtype avian influenza virus antigen by large-scale suspension culture in bioreactor.