{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T12:53:44Z","timestamp":1791550424476,"version":"4.3.4"},"reference-count":51,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T00:00:00Z","timestamp":1791331200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Key R&D Program of Ningxia Province","award":["2023BCF01013"],"award-info":[{"award-number":["2023BCF01013"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32373130"],"award-info":[{"award-number":["32373130"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["32373130"],"award-info":[{"award-number":["32373130"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology"],"abstract":"<jats:p>Rhinogobio nasutus, a freshwater fish belonging to the family Cyprinidae, is primarily distributed in the upper and middle reaches of the Yellow River. However, overfishing and habitat degradation have resulted in a marked decline in its wild populations, and the species was listed as a Class II nationally protected aquatic wildlife species in China in 2021. Spermatogonial stem cells (SSCs) provide a valuable resource for fish germplasm conservation and reproductive biotechnology. In this study, we successfully established a long-term cultured germ-cell population with spermatogonial stem cell-like characteristics derived from R. nasutus (RnSSCs). These cells exhibited a morphology consistent with an undifferentiated spermatogonial phenotype and showed a diploid chromosome number of 2n = 50. Alkaline phosphatase (AP) staining showed strong positive activity, while RT-PCR detected the expression of germ-cell- and stem-cell-associated markers, with only weak expression of the somatic-cell-associated marker. Immunofluorescence staining revealed strong signals for both proliferating cell nuclear antigen (PCNA) and the germ cell-specific protein Vasa. After more than four months of in vitro culture, the cells continued to proliferate and retained expression of germ-cell- and stem-cell-associated markers. Following transplantation into zebrafish blastulae, fluorescently labelled cells were detected in the gonadal region of a small number of recipient embryos. Following hormonal induction, some cells developed a sperm-like morphology, accompanied by the expression of the meiotic-associated markers rec8 and dmc1. The establishment of the RnSSC line provides a renewable cellular resource for studying spermatogenesis and germ cell transplantation in R. nasutus. These findings provide a useful cellular resource for studying the reproductive biology of R. nasutus.<\/jats:p>","DOI":"10.3390\/biology15191779","type":"journal-article","created":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T08:34:25Z","timestamp":1791448465000},"page":"1779","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Establishment and Characterization of a Spermatogonial Stem Cell-like Line with Potential for Meiotic Differentiation in the Endangered Fish Rhinogobio nasutus"],"prefix":"10.3390","volume":"15","author":[{"given":"Yue","family":"Wang","sequence":"first","affiliation":[{"name":"Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, College of Fisheries and Life Science, Shanghai Ocean University, Ministry of Education, Shanghai 201306, 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Shanghai Ocean University, Ministry of Education, Shanghai 201306, China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9404-3110","authenticated-orcid":false,"given":"Ni","family":"Hong","sequence":"additional","affiliation":[{"name":"Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, College of Fisheries and Life Science, Shanghai Ocean University, Ministry of Education, Shanghai 201306, 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China"}],"role":[{"vocabulary":"crossref","role":"author"},{"role":"conceptualization","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8729-7964","authenticated-orcid":false,"given":"Mingyou","family":"Li","sequence":"additional","affiliation":[{"name":"Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, College of Fisheries and Life Science, Shanghai Ocean University, Ministry of Education, Shanghai 201306, 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