Establishment and Characterization of a Spermatogonial Stem Cell-like Line with Potential for Meiotic Differentiation in the Endangered Fish Rhinogobio nasutus.
- 1Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, College of Fisheries and Life Science, Shanghai Ocean University, Ministry of Education, Shanghai 201306, China.
- 2Ningxia Fisheries Research Institute, Yinchuan 750001, China.
- 3Ningxia Fishery Engineering Technology Research Center, Yinchuan 750001, China.
Abstract
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.
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BibTeX
@article{wang2026establishment,
title = {{Establishment and Characterization of a Spermatogonial Stem Cell-like Line with Potential for Meiotic Differentiation in the Endangered Fish Rhinogobio nasutus.}},
author = {Yue Wang and Yanbin Liu and Muhammad Jawad and Haijing Xu and Qinjun Wu and Lang Gui and Ni Hong and Zongqiang Lian and Mingyou Li},
journal = {Biology},
year = {2026},
month = oct,
doi = {10.3390/biology15191779},
url = {https://europepmc.org/article/MED/42883452},
note = {CC BY},
}
RIS
TY - JOUR
TI - Establishment and Characterization of a Spermatogonial Stem Cell-like Line with Potential for Meiotic Differentiation in the Endangered Fish Rhinogobio nasutus.
AU - Yue Wang
AU - Yanbin Liu
AU - Muhammad Jawad
AU - Haijing Xu
AU - Qinjun Wu
AU - Lang Gui
AU - Ni Hong
AU - Zongqiang Lian
AU - Mingyou Li
PY - 2026
DA - 2026/10/07
JO - Biology
DO - 10.3390/biology15191779
UR - https://europepmc.org/article/MED/42883452
AB - 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.
SN - 2079-7737
ER -