Mammalian cell cultivation using nutrients extracted from microalgae

Abstract

Mammalian cells have been used in various research fields. More recently, cultured cells have been used as the cell source of “cultured meat.” Cell cultivation requires media containing nutrients, of which glucose and amino acids are the essential ones. These nutrients are generally derived from grains or heterotrophic microorganisms, which also require various nutrients derived from grains. Grain culture, in turn, requires many chemical fertilizers and agrochemicals, which can cause greenhouse gas emission and environmental contamination. Furthermore, grain production is greatly influenced by environmental changes. In contrast, microalgae efficiently synthesize various nutrients using solar energy, water, and inorganic substances, which are widely used in the energy sector. In this study, we aimed to apply nutrients extracted from microalgae in the culture media for mammalian cell cultivation. Glucose was efficiently extracted from Chlorococcum littorale or Arthrospira platensis using sulfuric acid, whereas 18 of the 20 proteinogenic amino acids were efficiently extracted from Chlorella vulgaris using hydrochloric acid. We further investigated whether nutrients present in the algal extracts could be used in mammalian cell cultivation. Although almost all C2C12 mouse myoblasts died during cultivation in a glucose‐ and amino acid‐free medium, the cell death was rescued by adding algal extract(s) into the nutrient‐deficient media. This indicates that nutrients present in algal extracts can be used for mammalian cell cultivation. This study is the first step toward the establishment of a new cell culture system that can reduce environmental loads and remain unaffected by the impact of environmental changes.

Formation of contractile 3D bovine muscle tissue for construction of millimetre-thick cultured steak

“…27 As the motility of myocytes in the gel is related to the rate of myotube formation, hydrogels with viscoelasticity similar to the fibrin-matrigel mixture would be essential for the edible alternative gel. Furthermore, although we use the animal serum at the present stage, it is necessary to culture the tissue using serum-free medium 28 or a medium from algae culture 29 for consumer acceptance and cost reduction.…”

In vitro culture of bovine fibroblasts using select serum-free media supplemented withChlorella vulgarisextract

“…Given that a common and relatively abundant signal feature found in the mass spectra of algae extract samples mapped to NAD + /NADP + as the parent species, it is possible that enriched levels of NAD(H)/NADP(H) present in the algae extracts may have caused enhanced activity of the dehydrogenase enzymes in the cells thus explaining the observed increase in viability. Further, it was previously shown that CVE also contains high levels of glutamate (Okamoto et al, 2020), as Chlorella algae use NADPH-dependent glutamate dehydrogenase enzymes to control their cell cycle (Talley et al, 1972). Glutamate has been shown to increase oxidation of NADH (Kannurpatti and Joshi, 1999; Maalouf et al, 2007), and increase cancer cell viability and growth (Gelb et al, 2015; Reiman et al, 1981; Takano et al, 2001; Yamaguchi et al, 2020).…”

“…Ng and colleagues report that extract derived from Chlorella vulgaris promoted growth of Chinese hamster ovary (CHO) and mesenchymal stem cells (MSC) under certain conditions, some of which did not contain any serum. In another study, researchers reported high levels of glucose and amino acids present in various microalgae extracts, with particularly high levels of aspartate and glutamate detected in extract from C. vulgaris (Okamoto et al, 2020); these authors proceeded to show that they could use such algae extracts as a substitute to culture cells in media deficient in glucose and amino acids. Here we tested the efficacy of culturing cells by replacing serum in the media with algae extract and found that we could achieve appreciable growth and viability of bovine fibroblasts using select serum-free media supplemented with C. vulgaris extract (CVE).…”

Simple and effective serum-free medium for sustained expansion of bovine satellite cells for cell cultured meat

“…It should be noted, however, that significant cost reduction of serum-free media is still needed to make cultured meat production economically viable from an industrial perspective. Here, reducing the cost of albumin, growth factors and basal media (e.g., through the use of plant or algal hydrolysates) will be essential 26,27 . Additionally, co-culture of meat-relevant cells with nutrient-or growth factor-producing cells could offer valuable cost-saving opportunities 28 .…”