国产与进口牛血清促进Vero细胞生长的比较

 目的  通过对国产与进口牛血清促进Vero细胞生长的比较，筛选可替代进口血清用于轮状病毒疫苗生产中Vero细胞培养的国产牛血清。方法  以国产的3批胎牛血清和1批新生牛血清为待评血清，以进口胎牛血清为对照血清，制备细胞培养液。在T175细胞培养瓶（T瓶）和2层细胞工厂中连续传代培养Vero细胞各3代，观察每一代次的培养上清液、细胞形态和细胞汇合度，计算细胞收获量和与对照血清相比较的相对增长率。采用单样本t检验对细胞收获量与生产要求的细胞产量（T瓶：>1.50×10⁷ 个/ml；细胞工厂：>3.00×10⁸ 个/ml）进行比较。根据细胞相对增长率判断国产血清与进口对照血清促细胞生长活性的相近程度。结果  国产血清培养的Vero细胞生长状态均良好。T瓶培养时，国产血清组的细胞收获量为（1.56～9.30）×10⁷ 个/ml，与生产要求细胞产量的差异有统计学意义（t=2.44～3.76，P<0.05），且t值均>0，说明符合生产要求。细胞相对增长率接近或超过100%。细胞工厂培养时，国产血清组的细胞收获量为（1.80～4.92）×10⁸ 个/ml，与生产要求产量的差异无统计学意义（t=－0.23～1.16，P>0.05），但是只有1批胎牛血清和新生牛血清的细胞收获量均值>3.00×10⁸ 个/ml，且细胞相对增长率>90%。 结论  经与进口牛血清比较，国产的1批胎牛血清和1批新生牛血清可替代进口血清，用于轮状病毒疫苗生产中的Vero细胞培养。

Comparison of domestic and imported bovine sera in promoting Vero cell growth

Objective  To screen domestic bovine serum for replacement of imported serum used for Vero cell culture in rotavirus vaccine production through comparing the cell growth promoted by domestic and imported bovine sera.  Methods  Three batches domestic fetal bovine sera and one batch domestic new-born calf serum were evaluated. One batch imported fetal bovine serum was used as control. The cell culture medium was separately prepared with the above bovine sera. Vero cells were sequentially cultured in T175 flask and 2-layer cell factory, 3 passages each. Culture supernatant, cell morphology, and cell confluence of each passage were observed. Cell yield and relative growth rate compared to the imported serum were calculated. The cell yield was compared with the required cell output (T flask: >1.50×10⁷ cells/ml; cell factory: >3.00×10⁸ cells/ml) by single-sample t-test. The approximation degree of cell growth activity promoted by the domestic and imported sera was judged according to the cell relative growth rate. Results  The growth status of all Vero cells cultured in domestic serum was good. When the cells were cultured in T flask, the range of cell yields was (1.56-9.30) ×10⁷ cells/ml, significantly different from the required cell output (t=2.44-3.76, P<0.05) , with t>0, indicating meeting the production requirement. The cell relative growth rates approached or exceeded 100%. When the cells were cultured in cell factory, there was no significant difference between the cell yield (1.80-4.92)×10⁸ cells/ml] in domestic serum group and the required cell output (t=－0.23-1.16, P>0.05). But the mean cell yields of only one batch fetal bovine serum and new-born calf serum were >3.00×10⁸ cells/ml, and their cell relative growth rates were >90%.  Conclusion  When compared with imported serum, one batch bovine serum and one batch new-born calf serum can replace the imported serum for the use of Vero cell culture in the rotavirus vaccine production.