Over the past 20 years, significant advances have been made in the understanding of cell and tissue culture in the fields of bioengineering, cell biology, and genetics; however, these achievements have largely focused on mammalian systems. In vitro studies on fish cells and tissues have been relatively limited, but the use of fish cell lines as in vitro models for environmental toxicology, particularly cytotoxicity analysis, has been significant. In addition, cultured cells have been used to study fish parasites and as in vitro models for screening immunostimulants. Skeletal muscle is the most important edible tissue in fish and rapid muscle growth determines the advantages of in vivo aquaculture. The generation of fish muscle cell lines can help to provide a reliable platform for deciphering the mechanism of fish skeletal muscle growth both in vivo and in vitro. In addition, cultured fish meat is a promising technology for animal protein-based foods and the concept of cell biomass meat from fish needs to be further developed. Our data demonstrated the feasibility of obtaining and culturing sterlet muscle cells in vitro. This study highlights the potential advantages of cell aquaculture over traditional fishing and aquaculture, and the potential applications of fish muscle cell lines in the study of fish skeletal muscle growth and the production of edible cultured fish meat products.