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作为一种血清替代物提取,可增强哺乳动物细胞生长和蛋白质表达。

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

作者信息

Ng Jian Yao, Chua Mei Ling, Zhang Chi, Hong Shiqi, Kumar Yogesh, Gokhale Rajeev, Ee Pui Lai Rachel

机构信息

Department of Pharmacy, Faculty of Science, National University of Singapore, Singapore, Singapore.

Roquette Innovation Center, Helios, Singapore, Singapore.

出版信息

Front Bioeng Biotechnol. 2020 Sep 15;8:564667. doi: 10.3389/fbioe.2020.564667. eCollection 2020.

Abstract

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 , 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.

摘要

由于基于抗体和基于细胞的疗法的迅速发展,全球细胞培养市场正在经历显著增长。这两种疗法都依赖于不同的“生物工厂”的生产能力,即原核细胞和真核细胞、植物或动物,以进行可靠的大规模生产。提高产量的能力是一个重要的关注点。在众多采用的策略中,通过培养基成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是经常进行的操作。最近也在探索向培养基中添加特定成分以调节宿主细胞的代谢。在本研究中,我们研究了提取的微藻生物活性成分,即小球藻生长因子(CGF),作为一种用于血清替代和诱导蛋白质表达的细胞培养添加剂的用途。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的化学指纹图谱,并评估了其增强哺乳动物宿主细胞增殖的能力。在降低血清条件下,CGF在二维和三维细胞培养中均成功促进了中国仓鼠卵巢细胞(CHO)和间充质干细胞(MSC)的生长,长达21天。此外,CGF保留了细胞功能,这在CHO细胞中蛋白质表达增加以及MSC中干细胞表型的维持中得到了体现。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中有广泛的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcc3/7522799/57018b8ffd31/fbioe-08-564667-g001.jpg

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