Data_Sheet_1_Chlorella vulgaris Extract as a Serum Replacement That Enhances Mammalian Cell Growth and Protein Expression.docx

The global cell culture market is experiencing significant growth due to the rapid advancement in antibody-based and cell-based therapies. Both rely on the capacity of different living factories, namely prokaryotic and eukaryotic cells, plants or animals for reliable and mass production. The ability to improve production yield is of important concern. Among many strategies pursued, optimizing the complex nutritional requirements for cell growth and protein production has been frequently performed via culture media component titration and serum replacement. The addition of specific ingredients into culture media to modulate host cells’ metabolism has also recently been explored. In this study, we examined the use of extracted bioactive components of the microalgae Chlorella vulgaris, termed chlorella growth factor (CGF), as a cell culture additive for serum replacement and protein expression induction. We first established a chemical fingerprint of CGF using ultraviolet-visible spectroscopy and liquid chromatography-mass spectrometry and evaluated its ability to enhance cell proliferation in mammalian host cells. CGF successfully promoted the growth of Chinese hamster ovary (CHO) and mesenchymal stem cells (MSC), in both 2D and 3D cell cultures under reduced serum conditions for up to 21 days. In addition, CGF preserved cell functions as evident by an increase in protein expression in CHO cells and the maintenance of stem cell phenotype in MSC. Taken together, our results suggest that CGF is a viable culture media additive and growth matrix component, with wide ranging applications in biotechnology and tissue engineering.

全球细胞培养市场正因基于抗体和细胞的治疗方法的迅速发展而经历显著增长。这两种治疗方法均依赖于不同生物工厂——即原核和真核细胞、植物或动物——的可靠和大规模生产能力。提高生产效率的能力备受关注。在众多追求的策略中，通过培养基组分滴定和血清替代来优化细胞生长和蛋白质生产的复杂营养需求已被频繁实施。近年来，向培养基中添加特定成分以调节宿主细胞代谢的研究亦被探索。在本研究中，我们考察了微藻小球藻（Chlorella vulgaris）提取的生物活性成分，称为小球藻生长因子（CGF），作为血清替代和蛋白质表达诱导的细胞培养添加剂的应用。我们首先使用紫外-可见光谱和液相色谱-质谱联用技术建立了CGF的化学指纹，并评估了其在哺乳动物宿主细胞中增强细胞增殖的能力。CGF成功促进了中国仓鼠卵巢（CHO）和间充质干细胞（MSC）在低血清条件下2D和3D细胞培养中长达21天内的生长。此外，CGF通过在CHO细胞中蛋白质表达的增加和在MSC中干细胞表型的维持来保留细胞功能。综上所述，我们的结果表明CGF是一种可行的培养基添加剂和生长基质成分，在生物技术和组织工程领域具有广泛的应用前景。