Development and characterization of red seabream (Pagrus major) muscle satellite cell lines for cultivated seafood: highlighting serum reduction and microcarrier expansion.

Cellular aquaculture requires well-characterized marine cell lines for sustainable seafood production. This study developed muscle satellite cell lines from red seabream (Pagrus major) (PMMSC) to support cultured seafood production. Primary cells, isolated via collagenase digestion (2.61 × 10⁵ cells/mL), were cultured in L- 15 medium under varying FBS levels (10-20%) and temperatures (20-28 °C). Optimal growth occurred at 24 °C with 10% FBS, forming dense monolayers by day 9. Differentiation into myotubes, induced with 2% horse serum, was confirmed by myogenic protein expression (Pax7, MYH, MyoD, MyoG, Desmin) over 12 d. PMMSC proliferation was evaluated under reduced-serum conditions supplemented with Spirulina platensis extract, an algae-derived alternative to fetal bovine serum. The algae extract improved cell proliferation, demonstrating its potential for sustainable seafood production. Additionally, 3D cell expansion using gelatin microcarriers in a bioreactor resulted in an 8.18-fold increase in cell count, showcasing its potential for scalable production. This research provides a valuable resource for the cellular aquaculture field by establishing a well-characterized red seabream muscle satellite cell line and demonstrating the potential of algae-based serum replacements.