Rational design of medium supplementation strategy for improved influenza viruses production based on analyzing nutritional requirements of MDCK Cells.
Rational design of medium supplementation strategy for improved influenza viruses production based on analyzing nutritional requirements of MDCK Cells.
复制标题
基于MDCK细胞营养需求分析，合理设计提高流感病毒产量的培养基补充策略。
DOI：
10.1016/j.vaccine.2014.10.067
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发表时间：
2014-12
期刊：
影响因子：
5.5
通讯作者：
Chen, Ze
中科院分区：
文献类型：
--
作者：
Ye, Zhaoyang;Tan, Wen-Song;Luo, Jian;Chen, Ze
使用细胞培养技术生产流感疫苗如今已变得流行。然而，要满足日益增长的流感疫苗需求，提高流感病毒在细胞中的产量是前提。为了实现这一目标，在本研究中，MDCK细胞在病毒生产过程中的营养需求进行了分析，并制定了相应的营养补料策略。根据消耗速率和相应的浓度优化，在感染时向维持培养基中补充葡萄糖和快代谢氨基酸。与未补充营养的培养物相比，感染后0-48 h期间的平均细胞特异性死亡率为0.013 h-1，在补充营养的培养物中降低了40.91%。测定了总病毒滴度、HA抗原蛋白浓度和细胞特异性病毒产量。（1.88 ± 0.23）× 103 HA单位/50 μL，11.70 ± 0.22 μg/mL和（10.06 ± 1.16）× 103个/细胞，分别比对照组高84.04 ± 22.50%、31.46 ± 2.87%和86.64 ± 25.81%。这些数据表明，在病毒生产过程中适当补充营养物质可以减少细胞死亡，提高细胞特异性病毒产量和流感病毒总产量。本研究为流感疫苗细胞培养技术的发展奠定了基础。
Influenza vaccine production using cell culture technology has become popular nowadays. However, to meet the ever increasing demand of influenza vaccine, it is prerequisite to improve the yield of influenza virus in cells. To achieve this, in the present study, the nutritional requirements of MDCK cells in the virus production process were analyzed and a nutrient-feeding strategy was developed accordingly. Based on the consumption rates and corresponding concentration optimization, glucose and fast metabolized amino acids were supplemented into the maintaining medium at the time of infection. Compared with the non-supplemented culture, the average cell specific death rate during 0–48 h post-infection was 0.013 h−1, which was 40.91% lower in the nutrient-supplemented culture. Total virus titer, HA antigen protein concentration and cell-specific virus yield were (1.88 ± 0.23) × 103HA units/50 μL, 11.70 ± 0.22 μg/mL and (10.06 ± 1.16) × 103virions/cell, respectively, which were 84.04 ± 22.50%, 31.46 ± 2.87% and 86.64 ± 25.81% higher than those in the control, respectively. These data showed that the appropriate supplementation of nutrients during virus production process could reduce cell death, and improve cell-specific virus yield and total influenza virus output. This study laid foundation for the development of cell culture technology for influenza vaccine production.
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