Lora Tubić

Maintaining sustainable production of healthy and nutritious food represents a major global challenge. Cultivated meat could reduce the environmental impact of livestock production, but it remains costly, largely due to the expense of fetal bovine serum, a common cell culture media supplement. Algal and cyanobacterial proteins, particularly phycobiliproteins, represent promising sustainable sources of potentially bioactive peptides that may contribute to the development of serum-reduced or serum-free culture media. This study investigated the potential of hydrolysates prepared using the protein extracts of the red alga Porphyra haitanensis and the cyanobacterium Arthrospira platensis as functional supplements for cell cultivation in a low-serum environment. Twelve hydrolysates were generated by treating protein extracts from Arthrospira platensis and Porphyra haitanensis with six proteolytic enzymes: pepsin, trypsin, chymotrypsin, subtilisin, thermolysin and proteinase K. The effects of these hydrolysates were studied in three cell models of mammalian (CHO-K1), avian (QM7), and fish (MACK1) origin. The effects of hydrolysates on cell viability, proliferation, and antioxidant activity were evaluated. A. platensis–chymotrypsin hydrolysate significantly improved QM7 cell viability compared to control in both 80% reduced-serum and tryptose phosphate broth conditions (p ≤ 0.0001). Additionally, peptide composition analysis and identification of amino acid sequences provided preliminary insights into potentially bioactive chromopeptides associated with the maintenance and promotion of cellular growth. Algae/cyanobacteria-derived hydrolysates positively regulated cell viability and proliferation in serum-reduced conditions, although effects were dependent on the algal species, enzymatic treatment, and cell type. These findings highlight their potential as sustainable supplements for alternative cell culture media.

Cultured meat technology is a form of cellular agriculture where meat is produced from animal cells grown in a lab, instead of raising and slaughtering animals. This technology relies heavily on fetal bovine serum (FBS) in cell media; hence, production is costly and contributes significantly to ammonia and greenhouse gas emissions. Achieving the successful commercialization of cell-cultured food requires the critical resolution of manufacturing cost and safety concerns. Hence, our research efforts are focused on identifying commercially viable and ecologically sustainable alternatives to FBS. In this study, we evaluated the potential of twenty-six water-based algal and cyanobacterial extracts to stimulate cell growth for meat cultivation under 90% reduced serum conditions. The extracts were compared in viability, proliferation, and Trypan blue exclusion assays. In the first screening phase, the extracts were evaluated in a ZEM2S (zebrafish) cell culture in a 1% FBS regimen. Based on their ability to exhibit protein tolerance or promote cell proliferation, ten extracts were selected and further assayed in a QM7 cell culture. The QM7 cell line (myoblasts from Japanese quail) is highly relevant for meat cultivation because of its ability to differentiate into muscle fibers. Extracts derived from two microalgae species, Arthrospira platensis (Spirulina) and Dunaliella tertiolecta, demonstrated the highest tolerance in cell culture, above 10 μg/mL (expressed as total protein concentration). Tolerance at a 100 μg/mL concentration was demonstrated exclusively using an extract of blue spirulina (commercially purified Spirulina), which supported cell growth through multiple passages.