文献类型：期刊文献

中文题名：流感病毒生产的维持培养基开发及细胞代谢特征分析

英文题名：Development of Maintenance Medium Using for Influenza Virus Production and Analysis of Cellular Metabolic Characteristics

作者：谢盼[1];叶倩[1];刘旭平[1];谭文松[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室

年份：2019

卷号：35

期号：5

起止页码：133

中文期刊名：生物技术通报

外文期刊名：Biotechnology Bulletin

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD2019_2020】；

基金：国家自然科学基金项目(201206040,21406066);国家科技重大专项(2013ZX10004003-003-003)

语种：中文

中文关键词：流感病毒;维持培养基;MDCK细胞;代谢

外文关键词：influenza virus;maintenance medium;MDCK cells;metabolism

摘要：采用Simplex Lattice设计开发出适于流感病毒在MDCK细胞中高效扩增的维持培养基,同时以细胞生长培养基作为对照组,研究该培养基下病毒感染后的细胞代谢特征。通过统计学方法,确定维持培养基由A和F培养基以0.586∶0.432比例混合组成,在2 L生物反应器中其所支持的最大病毒滴度为214.062 HAU/100μL,较对照组提高了59.55%。代谢分析表明,在葡萄糖初始浓度一致的条件下,维持培养基组增强了细胞对葡萄糖的摄取及糖酵解途径,对于利于病毒增殖的组分,如丝氨酸、精氨酸、异亮氨酸等,细胞的消耗速率显著增强;对于抑制病毒扩增的组分,如丙氨酸,细胞的生成速率大幅度下降。该研究方法和所得的关键细胞代谢特征对于其他病毒疫苗培养基的开发和优化提供了方法借鉴和理论指导。
This study developed a maintenance medium supporting the influenza virus production to high yield based on MDCK cell culture and using Simplex Lattice design,and studied the cellular metabolic characteristics during the virus infection under the maintenance medium,meanwhile the cell growth medium was as a control group. The maintenance medium was composed of A and F medium in a ratio of0.586∶0.432 by the statistical analysis,in which the maximum virus titer was 214.062 HAU/100 μL in the 2 L bioreactor,which was 59.55%higher than that of the control group. When the initial concentrations of glucose were the same,the cells in the maintenance medium group enhanced the glucose uptake and the glycolysis pathway,the consumption rate of serine,arginine and isoleucine that were conducive to virus proliferation significantly increased,and the production rate of alanine inhibiting virus proliferation greatly decreased,compared with the control group. Therefore,this work emphasized the importance of maintenance medium development;meanwhile the results provide a methodological reference and theoretical guidance for the development and optimization of other viral vaccine media.