3种细胞基质制备的麻疹血凝素效价的比较

目的比较3种细胞两种培养基制备的麻疹血凝素的效价。方法分别采用含10%新生牛血清的MEM培养基和含0.2%水解乳蛋白的199培养基培养原代人羊膜细胞、传代人羊膜细胞(FL)和Vero细胞,以0.01 MOI麻疹病毒L4株感染细胞,收获原制血凝素,采用Tween-80/乙醚处理,经血凝试验监测血凝素最佳收获时间,并测定原制血凝素和血凝素效价。结果采用MEM培养基培养的FL和Vero细胞在感染麻疹病毒后10~11 d,血凝素效价最高,均可达1∶16,原代人羊膜细胞在感染麻疹病毒后14~15 d,血凝素效价最高,可达1∶128;采用199培养基培养的FL和Vero细胞所制备的麻疹原制血凝素和血凝素效价均大于1∶32,高于MEM培养基组(1∶16),而原代人羊膜细胞所制备的麻疹原制血凝素和血凝素效价均为1∶128。结论 3种细胞两种培养基均可用于制备麻疹血凝素。

Titers of hemagglutinin of measles virus prepared with three cell substrates

Objective To compare the titers of hemagglutinin of measles virus prepared with three cell substrates in two media.Methods Primary human amniotic epithelial cells,FL cells and Vero cells were cultured in MEM containing 10% newborn bovine serum and 199 medium containing 0.2% lactalbumin hydrolysate separately,and infected with measles virus L4 strain at a MOI of 0.01.Plain hemagglutinin was harvested and treated with Tween-80 / ether to obtain hemagglutinin.The optimal time for harvesting hemagglutinin was monitored by hemagglutination test,and the plain hemagglutinin and hemagglutinin were determined for titer.Results The titers of hemagglutinin harvested 10 ~ 11 d after in fection of FL and Vero cells with measles virus in MEM reached the maximums of 1 ∶ 16.However,the titer of hemagglutinin harvested from human amniotic epithelial cells reached the maximum(1 ∶ 128)14 ~ 15 d after infection.Both the titers of plain hemagglutinin and hemagglutinin of measles virus infected to FL and Vero cells in 199 medium were more than 1︰32,which were higher than those in MEM(1 ∶ 16),while those infected to primary human amniotic epithelial cells in the two media were 1 ∶ 128.Conclusion All the three cell substrates and two media may be used for preparation of measles virus hemagglutinin.