Establishment and Characterization of a Long-Term Ovarian Cell Line (SBO) from Asian Seabass (Lates calcarifer) Expressing Germline Stem Cell Markers

Germline stem cells (GSCs) are crucial for gametogenesis, genetic conservation, and molecular breeding. Although GSCs lines have been well studied in mammals and several model teleost species, progress in commercial marine teleosts remains limited. In this study, we report a successful establishment of a long-term stable ovarian cell line derived from the Asian seabass (Lates calcarifer), designated the Seabass Ovarian (SBO) cell line. Ovaries were dissociated using a combined collagenase-trypsin digestion protocol; the cells were propagated and maintained in DMEM supplemented with bFGF, LIF, and fish serum. The SBO cells exhibited strong Alkaline Phosphatase activity. Furthermore, the cultured cells robustly expressed both germ-cell specific markers (Vasa) and pluripotency associated proteins (Nanog, SSEA-1). These findings indicate the successful isolation and long-term maintenance of an ovarian cell line expressing female germline stem cell markers from Asian seabass ovaries. The established cell line not only provides a valuable in vitro model for elucidating the mechanisms behind germ cell differentiation but also serves as a crucial cellular resource for advancing genetic breeding, germplasm preservation, and surrogate broodstock technologies in marine teleosts.