流感病毒生产的维持培养基开发及细胞代谢特征分析

doi:10.13560/j.cnki.biotech.bull.1985.2019-0056

生物技术通报 ›› 2019, Vol. 35 ›› Issue (5): 133-139.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0056

流感病毒生产的维持培养基开发及细胞代谢特征分析

谢盼, 叶倩, 刘旭平, 谭文松

华东理工大学生物反应器工程国家重点实验室,上海 200237

收稿日期:2019-01-19 出版日期:2019-05-26 发布日期:2019-05-23
作者简介:谢盼,女,硕士研究生,研究方向：动物细胞与组织工程;E-mail：xiepan1025@126.com
基金资助:
国家自然科学基金项目（201206040,21406066）,国家科技重大专项（2013ZX10004003-003-003）

Development of Maintenance Medium Using for Influenza Virus Production and Analysis of Cellular Metabolic Characteristics

XIE Pan, YE Qian, LIU Xu-ping, TAN Wen-song

State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai 200237

Received:2019-01-19 Published:2019-05-26 Online:2019-05-23

摘要/Abstract

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2^14.062 HAU/100 μL,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。

关键词: 流感病毒, 维持培养基, MDCK细胞, 代谢

Abstract: 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 of 0.586∶0.432 by the statistical analysis,in which the maximum virus titer was 2^14.062 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.

Key words: influenza virus, maintenance medium, MDCK cells, metabolism

谢盼, 叶倩, 刘旭平, 谭文松. 流感病毒生产的维持培养基开发及细胞代谢特征分析[J]. 生物技术通报, 2019, 35(5): 133-139.

XIE Pan, YE Qian, LIU Xu-ping, TAN Wen-song. Development of Maintenance Medium Using for Influenza Virus Production and Analysis of Cellular Metabolic Characteristics[J]. Biotechnology Bulletin, 2019, 35(5): 133-139.

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流感病毒生产的维持培养基开发及细胞代谢特征分析

Development of Maintenance Medium Using for Influenza Virus Production and Analysis of Cellular Metabolic Characteristics

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