H1N1流感病毒在微载体培养MDCK细胞上增殖的研究

Research on multiplication of the H1N1 subtype influenza virus in large-scale micro-carrier-based MDCK cell culture system

目的探索MDCK细胞在微载体上的培养条件,并研究H1N1型流感病毒在MDCK细胞上的增殖条件。方法在微载体上培养好MDCK细胞上用H1N1型流感病毒在不同的病毒感染复数(MOI)、胰酶浓度两个关键的病毒增殖条件进行流感病毒在细胞上的增殖研究。结果微载体质量浓度为6 g/L时,MDCK细胞培养密度可以达到4.5×106cells/mL。在MOI为0.05接种流感病毒,胰酶质量浓度4μg/mL,流感病毒在MDCK细胞上可获得较高的滴度。结论 MDCK细胞用微载体培养可以达到较高的细胞密度,可以作为规模化生产新型流感病毒疫苗的主要细胞基质进行进一步的研究。

Objective To investigate the growth kinetics of influenza virus subtype H1N1 in mammalian cell-based microcarrier culture system, and then optimize cultural conditions. Methods On the basis of diversity of MOI and TPCK-trypsin concentration and micro-cartier concentration, MDCK cells cultured in micro-carrier was infected by influenza virus. The density of MDCK cells and HA concentration were detected. Results At of 0.05 MOI and 4 μg/mL of TPCK-trypsin concentration, MDCK cell density was 4.5 ×106 cells/mL and HA concentration was higher with the concentration of microcarrier 6 g/L. Conclusion The results demonstrated that the influenza virus H1 N1 subtype could be grown in large-scale mammalian cell-based micro-carrier culture system with high-yield virus and evidenced the feasibility of mammalian cellbased micro-carrier culture system for influenza vaccine.