Continuous fish muscle cell line with capacity for myogenic and adipogenic-like phenotypes

• Published in: Scientific reports Vol. 13; no. 1; p. 5098
• Main Authors: Saad, Michael K., Yuen, John S. K., Joyce, Connor M., Li, Xinxin, Lim, Taehwan, Wolfson, Talia L., Wu, Justin, Laird, Jason, Vissapragada, Sanjana, Calkins, Olivia P., Ali, Adham, Kaplan, David L.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 29.03.2023; Nature Publishing Group; Nature Portfolio
• Subjects: 631/1647; 631/61; 631/80; Animals; Aquaculture; Biopsy; Cell culture; Cell Line; Commercial fishing; Fish; Fishes; Food safety; Genomes; Genotypes; Humanities and Social Sciences; Immortalization; Lipids; Mammalian cells; Mammals; Morphology; multidisciplinary; Muscle Cells; Muscles; Musculoskeletal system; Myosin; Perciformes - genetics; Phenotype; Phenotypes; Science; Science (multidisciplinary); Seafood; Skeletal muscle; Trout; Zebrafish
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-023-31822-2

Cell-cultivated fish offers the potential for a more ethical, sustainable, and safe seafood system. However, fish cell culture is relatively understudied in comparison to mammalian cells. Here, we established and characterized a continuous Atlantic mackerel ( Scomber scombrus ) skeletal muscle cell line (“Mack” cells). The cells were isolated from muscle biopsies of fresh-caught fish, with separate isolations performed from two distinct fish. Mack1 cells (cells from the first isolation) were cultured for over a year and subcultured over 130 times. The cells proliferated at initial doubling times of 63.9 h (± 19.1 SD). After a spontaneous immortalization crisis from passages 37–43, the cells proliferated at doubling times of 24.3 h (± 4.91 SD). A muscle phenotype was confirmed through characterization of muscle stemness and differentiation via paired-box protein 7 and myosin heavy chain immunostaining, respectively. An adipocyte-like phenotype was also demonstrated for the cells through lipid accumulation, confirmed via Oil Red O staining and quantification of neutral lipids. New qPCR primers ( HPRT , PAX3B , MYOD1 , MYOG , TNNT3A , and PPARG ) were tailored to the mackerel genome and used to characterize mackerel cell genotypes. This work provides the first spontaneously immortalized fish muscle cell line for research, ideally serving as a reference for subsequent investigation.