Development of oyster protein-enhanced scaffolds with seven-band grouper muscle satellite cells for cultured seafood production.

Abstract

Cell-based aquaculture holds significant promise for revolutionizing the seafood industry by addressing the limitations of traditional fishing and aquaculture practices. In this study muscle cells from Seven-band grouper (SBG) Epinephelus septemfasciatus were isolated using an enzymatic digestion method. Initial cell attachment and growth were monitored, showing robust proliferation when cultured in L-15 medium supplemented with 10% fetal bovine serum (FBS) and basic fibroblast growth factor (bFGF). We assessed the biocompatibility and cytotoxicity of two protein sources, oyster protein (OP) and soy protein (SP), for their effects on grouper muscle cell viability and growth. OP demonstrated strong biocompatibility, effectively supporting cell viability and significantly promoting muscle cell proliferation. At a concentration of 10,000 µg/mL, OP increased muscle cell proliferation by up to 90% after 48 h of incubation. SP, however, exhibited dose-dependent cytotoxic effects, with reduced cell viability observed at higher concentrations (10,000 µg/mL) over 48 h. This comparative analysis indicates that OP maintains cellular health and enhances cell growth, while SP may limit cell viability at elevated concentrations. Following these findings, we prepared a scaffold using Alginate-κ-Carrageenan (Alg-κ-Car) combined with oyster protein (Alg-κ-Car-OP), which showed enhanced gelation and printability properties. 3D bioprinting of grouper muscle satellite cells (GMSC) within Alg-κ-Car-OP scaffolds resulted in higher cell viability than Alg-κ-Car scaffolds alone. Taste sensory analysis using an electronic tongue revealed distinct taste profiles, with Alg-κ-Car-OP-GMSC scaffolds exhibiting the highest umami score. Flavor analysis using flash gas chromatography and an electronic nose differentiated between scaffold types and protein samples, highlighting potential flavor markers. These findings underscore the potential of cell-based aquaculture, especially with OP-incorporated scaffolds, to meet the demand for sustainable and nutritious seafood alternatives. Further research is warranted to optimize production processes and explore commercial applications.