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

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

doi:10.3389/fbioe.2020.564667

Rajeev Gokhale, Pui Lai Rachel Ee + Show 5 more

Open Access

https://doi.org/10.3389/fbioe.2020.564667

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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 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.

Highlights

Therapeutic proteins are an important class of medications for the treatment of complex diseases such as diabetes and immunological defects
We present the characterization of extracted bioactive components of C. vulgaris, termed chlorella growth factor (CGF), and demonstrate their ability to support cell growth in low serum conditions using high utility mammalian cell lines, such as Chinese hamster ovary (CHO) and mesenchymal stem cells (MSCs)
To establish the chemical fingerprint of CGF, phenolic compounds within CGF were separated by liquid chromatography, and the precursor ions present in each peak were subjected to collision-induced associated (CID) mass spectrometry analysis

Summary

INTRODUCTION

Therapeutic proteins are an important class of medications for the treatment of complex diseases such as diabetes and immunological defects. Mammalian cells have become the main host systems for recombinant protein production (Walsh, 2014) This is due to their innate membranous organelles allowing post-translational modification (PTM) of the expressed proteins, which is critical for certain proteins to fold properly and/or exert its biological activity (Amann et al, 2019). We present the characterization of extracted bioactive components of C. vulgaris, termed chlorella growth factor (CGF), and demonstrate their ability to support cell growth in low serum conditions using high utility mammalian cell lines, such as CHO and mesenchymal stem cells (MSCs). Our results present an exciting opportunity for commercial protein production, and for the development of novel MSC-based therapeutic strategies for tissue engineering and regenerative medicine (TERM)

MATERIALS AND METHODS

RESULTS AND DISCUSSION

Coli undetectable

CONCLUSION