文献类型：期刊文献

中文题名：悬浮MDCK-STAT1-KO工程细胞的流感病毒生产过程研究

英文题名：Influenza Virus Production Characteristics of Suspended MDCK-STAT1-KO Engineered Cells

作者：姚鸿[1];崔乃香[1];谭文松[1];赵亮[1];叶倩[1]

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

年份：2025

卷号：45

期号：5

起止页码：1

中文期刊名：中国生物工程杂志

外文期刊名：China Biotechnology

收录：;北大核心:【北大核心2023】；

语种：中文

中文关键词：流感病毒疫苗;MDCK工程细胞;高效生产

外文关键词：Influenza virus vaccine;MDCK engineered cell line;Effective production

摘要：目的:MDCK-STAT1-KO工程细胞已被证实可高效生产H1N1流感病毒,但受限于培养规模和血清依赖性等贴壁培养固有的性质,该细胞株难以应用到实际的疫苗生产中。单细胞悬浮培养易于放大,操作简便,备受疫苗生产青睐。但MDCK-STAT1-KO工程细胞在悬浮培养条件下的病毒生产情况及其病毒感染应激特征尚不清楚,限制了其进一步应用。方法:采用生产上的常规培养方式(传代培养和批式培养)和接毒条件[感染复数(MOI)=0.001],考察悬浮MDCK-STAT1-KO工程细胞(susMDCK-STAT1^(-))的生长和病毒生产情况;改变关键接毒参数,考察MOI、营养条件和毒株类型对susMDCK-STAT1^(-)细胞产毒的影响。设置同步感染实验,考察该细胞在病毒感染后的细胞凋亡水平和细胞免疫应激水平。结果:susMDCK-STAT1^(-)细胞生产H1N1病毒的最大HA滴度和最大Svy分别为(12.40±0.11)log2HAU/100μL和(15.86±1.34)×10^(3) virions/cell,均为H1N1病毒生产较高水平,同时改变关键工艺参数均不会影响该细胞株高效生产流感病毒的特性。此外,susMDCK-STAT1^(-)细胞具有一定抗凋亡特性,以及其胞内全局性的低免疫防御水平,有利于病毒快速扩增。结论:susMDCK-STAT1^(-)细胞可高效生产流感病毒,同时其抗凋亡特性及胞内低免疫水平是该细胞株高效生产的关键因素。该细胞有望未来应用于流感病毒疫苗的实际生产中,提高流感病毒生产水平。
Objective:The MDCK-STAT1-KO engineered cell line has been shown to efficiently produce H1N1 influenza virus;however,its application in influenza vaccine production is limited by the inherent characteristics of adherent culture,such as scale-up challenges and serum dependence.Single cell suspension culture is preferred for vaccine production due to its ease of scale-up and operation.Nevertheless,the viral production capabilities and stress response characteristics of the MDCK-STAT1-KO cell line under suspension culture conditions remain unclear,which restricts its further application and research.Methods:The growth characteristics of suspension MDCK-STAT1-KO cells(susMDCK-STAT1^(-))were assessed using both passage and batch culture methods.The viral production capacity of these cells was assessed under conventional infection conditions(MOI=0.001).Key infection parameters,including MOI,nutrient conditions,and viral strain type,were modified to evaluate their effects on viral yield from susMDCK-STAT1^(-)cells.Synchronous infection experiments were performed to investigate cell apoptosis levels and immune stress responses following viral infection.Results:The maximum hemagglutinin(HA)titer and maximum specific virus yield(Svy)produced by susMDCK-STAT1^(-)cells were(12.40±0.11)log2HAU/100μL and(15.86±1.34)×10^(3) virions/cell,respectively,indicating a high production level for H1N1 virus.Moreover,alterations in key process parameters did not affect the cell line's ability to efficiently produce influenza virus.Additionally,susMDCK-STAT1^(-)cells exhibited certain anti-apoptotic properties and an overall lower level of immune defense,compared to the wild-type MDCK suspension cell line(susMDCK),facilitating virus invasion and replication and rapid virus release.Conclusions:susMDCK-STAT1^(-)cells are capable of efficiently producing influenza virus,and their anti-apoptotic properties and low intracellular immune response levels are critical factors contributing to the highly efficient virus production process.This cell line shows promise for future applications in the practical production of influenza virus vaccines,enhancing the overall production levels of influenza viruses.