大豆蛋白水解物促进MDCK细胞生长及流感病毒增殖的实验研究

Effect of soy protein hydrolysate on MDCK cell growth and influenza virus proliferation

目的 研究大豆蛋白水解物对Madin-Darby犬肾（Madin-Darby canine kidney，MDCK）细胞生长和流感病毒增殖的促进作用。方法 比较MDCK细胞在10%进口胎牛血清（fetal bovine serum，FBS）-Dulbecco改良Eagle培养基（Dulbecco’s modified Eagle’s medium，DMEM），10%国产新生牛血清（newborn calf serum，NCS）-DMEM和添加2、4、8 g/L大豆蛋白水解物SHEFF-VAX PLUS PF ACF（PF ACF）的10% 国产NCS -DMEM中的生长情况，以及甲型H7N9流感病毒接种细胞后的血凝素（hemagglutinin，HA）效价。结果 采用培养瓶静置培养时，5种培养体系中的细胞数在第3天达到最高，分别为（8.67±1.54）×106、（4.78±0.64）×106、（10.60±0.23）×106、（9.17±0.51）×106和（4.79±0.19）×106。采用搅拌瓶内微载体培养时，FBS-DMEM、NCS-DMEM和添加2、4 g/L PF ACF的NCS-DMEM中的细胞密度最高可达到（2.56±0.68）×106、（1.74±1.77）×106、（2.20±1.48）×106和（2.50±1.25）×106细胞/ml。流感病毒接种细胞后，FBS-DMEM、NCS-DMEM和添加4 g/L PF ACF的NCS-DMEM中测得的病毒HA效价分别为8.0、3.5和8.0 log2HAU/50 μl。结论 含国产NCS的培养基在添加4 g/L PF ACF后，MDCK细胞密度和流感病毒产量与含进口FBS的培养基的培养结果相仿，因此，大豆蛋白水解物可作为培养基添加物应用于MDCK细胞培养和流感病毒增殖。

To study the improvement by soy protein hydrolysate on Madin-Darby canine kidney （MDCK） cell growth and influenza virus replication. Methods MDCK cells were cultured in 10% imported fetal bovine serum（FBS）-Dulbecco’s modified Eagle’s medium （DMEM）, 10% domestic newborn calf serum （NCS）-DMEM and 10% NCS-DMEM supplemented with 2, 4, 8 g/L soy protein hydrolysate SHEFF-VAX PLUS PF ACF （PF ACF）, respectively. The growth of MDCK cells in different media and hemagglutinin （HA） titers after H7N9 influenza virus inoculation were compared. Results The maximum numbers of MDCK cells statically cultured with five culture systems in culture flask on day 3 reached （8.67±1.54）×106, （4.78±0.64）×106, （10.60±0.23）×106, （9.17±0.51）×106 and （4.79±0.19）×106, respectively. The highest densities of MDCK cells on microcarrier in spinner flask were （2.56±0.68）×106, （1.74±1.77）×106, （2.20±1.48）×106 and （2.50±1.25）×106 cells/ml when cultured in FBS-DMEM, NCS-DMEM and NCS-DMEM supplemented with 2, 4 g/L PF ACF, respectively. The HA titers of H7N9 virus replicated in FBS-DMEM, NCS-DMEM and NCS-DMEM supplemented with 4 g/L PF ACF were 8.0, 3.5 and 8.0 log2HAU/50 μl. Conclusions Adding 4 g/L PF ACF to domestic NCS-containing medium could enhance cell density and HA titer equivalent to imported FBS-containing medium. Thus soy protein hydrolysate can be used as a medium supplement for MDCK cell culture and influenza