Abstract：
Chinese hamster ovary (CHO) cells represent the most preferential host cell system for therapeutic monoclonal antibody (mAb) production. Enhancing mAb production in CHO cells can be achieved by adding chemical compounds that regulate the cell cycle and cell survival pathways. This study investigated the impact of ectoine supplementation on mAb production in CHO cells. The results showed that adding ectoine at a concentration of 100 mM on the 3rd day of cultivation improved mAb production by improving cell viability and extending the culture duration. RNA sequencing analysis revealed differentially expressed genes associated with cell cycle regulation, cell proliferation, and cellular homeostasis, in particular promotion of cell cycle arrest, which was then confirmed by flow cytometry analysis. Ectoine-treated CHO cells exhibited an increase in the number of cells in the G0/G1 phase. In addition, the cell diameter was also increased. These findings support the hypothesis that ectoine enhances mAb production in CHO cells through mechanisms involving cell cycle arrest and cellular homeostasis. Overall, this study highlights the potential of ectoine as a promising supplementation strategy to enhance mAb production not only in CHO cells but also in other cell lines.
摘要：
中国仓鼠卵巢(CHO)细胞代表用于治疗性单克隆抗体(mAb)生产的最优先的宿主细胞系统。增强CHO细胞中的mAb产生可以通过添加调节细胞周期和细胞存活途径的化合物来实现。该研究调查了在CHO细胞中补充外托宁对mAb产生的影响。结果表明,在培养的第3天以100mM的浓度添加外托宁通过改善细胞活力和延长培养持续时间来改善mAb的产生。RNA测序分析揭示了与细胞周期调控相关的差异表达基因，细胞增殖，和细胞内稳态，特别是促进细胞周期停滞，然后通过流式细胞术分析证实。以替托因处理的CHO细胞表现出G0/G1期细胞数量的增加。此外,细胞直径也增加。这些发现支持了以下假设：通过涉及细胞周期停滞和细胞稳态的机制，etoine增强了CHO细胞中mAb的产生。总的来说,这项研究强调了etoine作为一种有希望的补充策略的潜力，不仅可以在CHO细胞中而且可以在其他细胞系中提高mAb的产量。
