文献类型：期刊文献

中文题名：猪流行性腹泻病毒Vero细胞微载体培养条件的优化试验

英文题名：Optimizing experiment of culture conditions of porcine epidemic diarrhea virus in Vero cells microcarrier

作者：王建超[1];周燕[1];华平[1];邹勇[2];谭文松[1];钱永清[2]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237;[2]上海市农业科学院畜牧兽医研究所,上海201106

年份：2004

卷号：34

期号：5

起止页码：50

中文期刊名：中国兽医科技

外文期刊名：Chinese Journal of Veterinary Science and Technology

收录：北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；

基金：上海市科技兴农重点攻关项目 [农科攻字(99) 5 1和农科攻字 (2 0 0 1) 3 8]

语种：中文

中文关键词：猪流行性腹泻病毒;Vero细胞;微载体;生物反应器;疫苗;细胞培养;猪流行性腹泻

外文关键词：Vero cells; porcine epidemic diarrhea virus;microcarrier;bioreactor

摘要：通过研究搅拌速度、pH值、血清浓度等环境条件对细胞贴壁的影响以及细胞接种密度、微载体浓度与细胞生长的关系 ,对应用微载体技术和生物反应器系统生产猪流行性腹泻病毒(PEDV )的可行性进行了探讨。结果表明 ,搅拌转速、pH值、血清浓度均对Vero(PEDV )细胞在CT 3微载体上的贴壁率和均匀分布有影响 ;当接种密度为 15 .0× 10 4 /mL时 ,即每 1mg微载体接种 5 .0× 10 4 细胞时 ,可以获得较高的细胞密度 ;随着微载体浓度的提高 ,最高细胞密度也增加 ;在CelliGen反应器中 ,采用优化的培养条件 ,细胞密度可达到 174 .0× 10 4 /mL ,是方瓶培养的 3.5倍 ,每个细胞的PEDV含量与方瓶培养相当 。
Production of porcine epidemic diarrhea virus(PEDV) in microcarrier and bioreactor system, and some factors influencing cells attachmentand growth on microcarriers were investigated in this experiment. It was found that agitation speed, pH and serum concentration affected ratio of cell attachment and distribution uniformity on microcarriers. Serial experiments on the relation between cell seeding densities, microcarrier concentrations, and the growth of Vero cells had shown that optimal cell growth was obtained when the cell seeding density was 15.0×10^(4 )cells/mL or 5.0×10^(4)cells/mg microcarrier. The maximum cell density during cultivation in spinner flasks increased with microcarrier concentration increasing. Importantly, the yield of PEDV per cell in a bioreactor system with microcarriers was similar to that in T-flasks, but the cell density reached in the bioreactor was found 3.5-fold higher than that in T-flasks. In addition, the controlled animal tests shown that PEDV vaccine produced in the bioreactor system had comparable (immunogenicity). In conclusion, the experimental results from this work demonstrated that the bioreactor system with microcarriers is potential for industrial production of the PEDV vaccine.