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Proliferation and differentiation of primary bovine myoblasts using Chlorella vulgaris extract for sustainable production of cultured meat
Authors:Yuta Okamoto  Yuji Haraguchi  Azumi Yoshida  Hironobu Takahashi  Kumiko Yamanaka  Naoya Sawamura  Toru Asahi  Tatsuya Shimizu
Affiliation:1. Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda University, Tokyo, Japan

Institute of Advanced Biomedical Engineering and Science, TWIns, Tokyo Women's Medical University, Tokyo, Japan

Contribution: Conceptualization (equal), Data curation (lead), Formal analysis (lead), ​Investigation (lead), Methodology (lead), Resources (lead), Software (lead), Validation (lead), Visualization (lead), Writing - original draft (lead), Writing - review & editing (lead);2. Institute of Advanced Biomedical Engineering and Science, TWIns, Tokyo Women's Medical University, Tokyo, Japan;3. Institute of Advanced Biomedical Engineering and Science, TWIns, Tokyo Women's Medical University, Tokyo, Japan

Contribution: Methodology (supporting), Resources (supporting), Visualization (supporting), Writing - review & editing (supporting);4. Institute of Advanced Biomedical Engineering and Science, TWIns, Tokyo Women's Medical University, Tokyo, Japan

Contribution: Resources (supporting), Writing - review & editing (supporting);5. Research Organization for Nano & Life Innovation, Waseda University, Tokyo, Japan;6. Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda University, Tokyo, Japan

Faculty of Science and Engineering, Waseda University, Tokyo, Japan

Contribution: Methodology (supporting), Supervision (equal), Writing - review & editing (supporting)

Abstract:Recently, cultured meat obtained from livestock-derived cells is being considered as a sustainable food source that reduces the use of natural resources. This study aimed to show that nutrients extracted from Chlorella vulgaris were beneficial in the culture of primary bovine myoblasts (PBMs), a major cell source for cultured meat production. Nutrients (glucose, amino acids, and vitamins) present in the animal-cell culture media were effectively recovered from C. vulgaris using acid hydrolysis treatment. On culture in nutrient-free inorganic salt solution, cell death was induced in most PBMs after 6 days of cultivation. However, the addition of C. vulgaris extract (CVE) significantly improved PBM viability, which was comparable to the viability in conventional culture medium (Dulbecco's modified Eagle's medium). Furthermore, by adding horse serum to induce differentiation, the formation of myotubes was confirmed when CVE were used. Together, the results showed that CVE could be used as an alternative to the conventional culture medium for PBMs. These findings will not only lower the environmental risks associated with the establishment of this eco-friendly cell culture system, but also highlight microalgae as a potent nutrient source that can replace conventional grain-dependent nutrient sources.
Keywords:Chlorella vulgaris  cultured meat  microalgal extract  myotube formation  nutrients  primary bovine myoblast
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