MDCK细胞悬浮驯化及对H9亚型禽流感病毒敏感性的研究

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

关键词:  MDCK悬浮细胞  禽流感病毒  敏感性

Key words:  MDCK suspension cell  influenza virus  sensitivity

摘要:

为了获得对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培养的MDCK细胞进行了的无血清纯悬浮驯化，并最终获得了一株对H9亚型禽流感敏感的纯悬浮的细胞株MDCK-sus。通过测定表明，该细胞株能够迅速增殖，倍增时间为24 h，使用H9亚型禽流感病毒BX13株感染该细胞，培养72 h，培养液的病毒HA价可达到1:512，每0.1 mL病毒含量达到108.5 EID50,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。

Abstract:

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.