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

摘要

[Objective]To screen the best culture media for proliferation of avian influenza virus ( AIV) H9 subtype in MDCK cells. [ Method] The MDCK monolayer cells were respectively cultured in DMEM containing 10% (V/V) newborn calf serum (NBS) ,low -serum culture medium (MEM - MD -611) and serum - free medium (SFE4Mega) and then inoculated with different dilutions of AIV H9 subtype. These three kinds of media containing 10 (Ag/ml trypsin were also used for maintenance culture of AIV. Every 24 h,cytopathic changes were observed and the HA titer of the culture supernatant was also determined. [Result] After 72 -96 h of culture,the HA titers of the culture supernatant derived from the serum - free medium, low - serum culture medium and serum containing medium were in a decreasing manner. [ Conclusion ] The DMEM containing 10% (V/V) newborn calf serum ( NBS),low - serum culture medium (MEM-MD-611) and serum-free medium (SFE4Mega) can be used for proliferation of AIV, and the latter two may be more suitable to prepare AIV culture.%[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10 μg/ml胰蛋白酶的3种培养基作为维持液,培养病毒,每隔24h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.