兽用生物制品生产用牛血清质量标准研究——牛血清对Sp2/0细胞增殖试验研究

Study of Bovine Serum for Veterinary Biologics Production——Efficacy of Facilitating Sp2/0 Proliferation

关键词:  牛血清  Sp2/0细胞  克隆形成率  质量标准

Key words:  bovine serum  Sp2/0  cloning forming efficiency  criteria

摘要:

为制定兽用生物制品生产用牛血清对Sp2/0细胞增殖试验的质量标准，将处于对数生长期的Sp2/0细胞分散，配制成50万/ml细胞悬液，用含10%不同牛血清的MEM营养液，在96孔细胞培养板上作对倍稀释，在5%CO2培养箱37℃培养48h，计数每种血清对Sp2/0细胞的绝对克隆形成率。结果，胎牛血清对Sp2/0细胞的绝对克隆形成率为20～26%，犊牛血清对Sp2/0细胞的绝对克隆形成率为12～18%。以胎牛血清作为标准参考血清计算不同牛血清对Sp2/0细胞的相对克隆形成率，3批胎牛血清的相对克隆形成率均在80%以上，6批有效期之内的新生牛血清的相对克隆形成率为50%左右。因此，将兽用生物制品生产用胎牛血清对Sp2/0细胞增殖试验的质量标准规定为对标准血清相对克隆形成率不低于80%；新生犊牛血清对Sp2/0细胞增殖试验的质量标准规定为对标准血清相对克隆形成率不低于50%。

Abstract:

To determine the criteria of different class bovine serum facilitating Sp2/0 proliferation in the veterinary biologics production, the Sp2/0 cells in the logarithmic phase were dispersed, counted and confected a suspension of 5×105cells/ml with MEM. After distributing into several bottles, add 10% different batches bovine serum to these bottles and inoculate the suspension to 96-well plates at 2×series respectively. After Incubating at 37℃, 5%CO2 for 48 hours, calculate cloning forming efficiency of each serum to Sp2/0. Results show, the absolute cloning forming efficiency of fetal bovine serum (FBS) is 20~26%, the newborn bovine serum (NBS) is 12~18%. Each relative cloning forming efficiency of 3 batches of FBS is more than 80%, the 6 batches of NBS is respectively about 50%. From the above results, we determined the following criteria, the relative cloning forming efficiency of FBS is not less than 80% and the relative cloning forming efficiency of NBS is not less than 50%.