Yu Shouzhi, Yan Junyu, Yang Zhaona, Zhao Yuxiu, Wang Hui, Yang Xiaoming
Beijing Institute of Biological Products Company Limited, Beijing 100176, China.
China National Biotec Group Company Limited, Beijing 100024, China.
Vaccines (Basel). 2022 Oct 25;10(11):1787. doi: 10.3390/vaccines10111787.
The production efficiency of a cell substrate directly affects the yield of target products such as viruses, while its density is mainly regulated by the type of culture medium and culture conditions. In this study, Vero cells were used as model cells for systematic medium screening, and a high-efficiency medium for biological drug production was identified. Through the results of cell proliferation by a cell counting kit (CCK)-8 assay, 5-Ethynyl-2'-deoxyuridine(EdU) assay, real-time quantitative PCR (RT-qPCR) and Western blotting, we found that adding an appropriate amount of vitamin B to the conventional basic medium can significantly improve and maintain the high-density growth of Vero cells. In addition, the molecular mechanism of the high-density culture of Vero cells promoted by B vitamins is explained for the first time by using the systems multi-omics analysis methods. Here, we determined that B vitamins regulate cell proliferation through the synthesis and metabolism of unsaturated fatty acids, affecting the productivity of cell substrate in industrial production. This study provides an important tool for the screening of key components of cell-based high-efficiency medium.
细胞底物的生产效率直接影响病毒等目标产物的产量，而其密度主要受培养基类型和培养条件的调控。在本研究中，将Vero细胞用作模型细胞进行系统的培养基筛选，并鉴定出一种用于生物药物生产的高效培养基。通过细胞计数试剂盒（CCK）-8法、5-乙炔基-2'-脱氧尿苷（EdU）法、实时定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测细胞增殖结果，我们发现向传统基础培养基中添加适量的维生素B可显著改善并维持Vero细胞的高密度生长。此外，首次运用系统多组学分析方法阐释了B族维生素促进Vero细胞高密度培养的分子机制。在此，我们确定B族维生素通过不饱和脂肪酸的合成与代谢调节细胞增殖，影响工业生产中细胞底物的生产力。本研究为基于细胞的高效培养基关键成分筛选提供了重要工具。
