Establishment and Characteristics of the Spermatogonial Stem Cell Line from the Yellow River Carp (<i>Cyprinus carpio haematopterus</i>)

To address the growing consumer demands for improved fish meat quality, desirable morphological traits, and sustainable production practices, researchers have intensified efforts in the selective breeding and genetic improvement of carp (<i>Cyprinus carpio</i>) varieties. However, tradit...

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Main Authors: Huijie Zhou, Tianqi Liu, Tan Zhang, Zhipeng Sun, Huan Xu, Tingting Zhang, Yashan Yin, Na Li, Ting Yan, Youyi Kuang
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Biology
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Online Access:https://www.mdpi.com/2079-7737/14/5/536
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author Huijie Zhou
Tianqi Liu
Tan Zhang
Zhipeng Sun
Huan Xu
Tingting Zhang
Yashan Yin
Na Li
Ting Yan
Youyi Kuang
author_facet Huijie Zhou
Tianqi Liu
Tan Zhang
Zhipeng Sun
Huan Xu
Tingting Zhang
Yashan Yin
Na Li
Ting Yan
Youyi Kuang
author_sort Huijie Zhou
collection DOAJ
description To address the growing consumer demands for improved fish meat quality, desirable morphological traits, and sustainable production practices, researchers have intensified efforts in the selective breeding and genetic improvement of carp (<i>Cyprinus carpio</i>) varieties. However, traditional breeding methods are often time-consuming and inefficient, which poses challenges to the sustainable development of the carp aquaculture industry. The establishment of germ stem cell lines offers a crucial tool for the study of germ cells, genetic improvement, and species conservation. In this study, we successfully established a spermatogonial stem cell line (YRSSCs) from Yellow River carp (<i>Cyprinus carpio haematopterus</i>) that can be cultured in vitro for the long term. We optimized the culture conditions to maintain their self-renewal and differentiation capabilities. The results demonstrated that YRSSCs have a diploid karyotype and can stably proliferate for over a year in L-15 medium supplemented with 5 mmol/L HEPES, 50 μmol/L β-mercaptoethanol, 15% FBS, 2 ng/mL bFGF, 2 ng/mL LIF, 1% carp serum, 800 IU/mL penicillin, 0.8 mg/mL streptomycin, 2 μg/mL amphotericin B, 1% zebrafish embryo extract, and 1% glutamine at 30 °C in the absence of CO<sub>2</sub>. The cells exhibited a typical germ stem cell gene expression profile, with strong expression of the <i>vasa</i>, <i>plzf-a</i>, and <i>Oct4-a</i> genes. Additionally, this study found that YRSSCs possess the ability to differentiate in vitro and functionally colonize in vivo within recipient bodies. This research explored the establishment of YRSSCs and their differentiation potential both in vitro and in vivo, providing a novel strategy for the genetic improvement of aquaculture fish species through germ stem cell-based gene editing and transplantation technologies.
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spelling doaj-art-696a1c4e4c25447a85ee15110de72b7a2025-08-20T01:56:17ZengMDPI AGBiology2079-77372025-05-0114553610.3390/biology14050536Establishment and Characteristics of the Spermatogonial Stem Cell Line from the Yellow River Carp (<i>Cyprinus carpio haematopterus</i>)Huijie Zhou0Tianqi Liu1Tan Zhang2Zhipeng Sun3Huan Xu4Tingting Zhang5Yashan Yin6Na Li7Ting Yan8Youyi Kuang9National and Local Joint Engineering Laboratory for Freshwater Fish Breeding, No. 232, Hesong Street, Daoli District, Harbin 150070, ChinaNational and Local Joint Engineering Laboratory for Freshwater Fish Breeding, No. 232, Hesong Street, Daoli District, Harbin 150070, ChinaNational and Local Joint Engineering Laboratory for Freshwater Fish Breeding, No. 232, Hesong Street, Daoli District, Harbin 150070, ChinaNational and Local Joint Engineering Laboratory for Freshwater Fish Breeding, No. 232, Hesong Street, Daoli District, Harbin 150070, ChinaNational and Local Joint Engineering Laboratory for Freshwater Fish Breeding, No. 232, Hesong Street, Daoli District, Harbin 150070, ChinaNational and Local Joint Engineering Laboratory for Freshwater Fish Breeding, No. 232, Hesong Street, Daoli District, Harbin 150070, ChinaNational and Local Joint Engineering Laboratory for Freshwater Fish Breeding, No. 232, Hesong Street, Daoli District, Harbin 150070, ChinaNational and Local Joint Engineering Laboratory for Freshwater Fish Breeding, No. 232, Hesong Street, Daoli District, Harbin 150070, ChinaNational and Local Joint Engineering Laboratory for Freshwater Fish Breeding, No. 232, Hesong Street, Daoli District, Harbin 150070, ChinaNational and Local Joint Engineering Laboratory for Freshwater Fish Breeding, No. 232, Hesong Street, Daoli District, Harbin 150070, ChinaTo address the growing consumer demands for improved fish meat quality, desirable morphological traits, and sustainable production practices, researchers have intensified efforts in the selective breeding and genetic improvement of carp (<i>Cyprinus carpio</i>) varieties. However, traditional breeding methods are often time-consuming and inefficient, which poses challenges to the sustainable development of the carp aquaculture industry. The establishment of germ stem cell lines offers a crucial tool for the study of germ cells, genetic improvement, and species conservation. In this study, we successfully established a spermatogonial stem cell line (YRSSCs) from Yellow River carp (<i>Cyprinus carpio haematopterus</i>) that can be cultured in vitro for the long term. We optimized the culture conditions to maintain their self-renewal and differentiation capabilities. The results demonstrated that YRSSCs have a diploid karyotype and can stably proliferate for over a year in L-15 medium supplemented with 5 mmol/L HEPES, 50 μmol/L β-mercaptoethanol, 15% FBS, 2 ng/mL bFGF, 2 ng/mL LIF, 1% carp serum, 800 IU/mL penicillin, 0.8 mg/mL streptomycin, 2 μg/mL amphotericin B, 1% zebrafish embryo extract, and 1% glutamine at 30 °C in the absence of CO<sub>2</sub>. The cells exhibited a typical germ stem cell gene expression profile, with strong expression of the <i>vasa</i>, <i>plzf-a</i>, and <i>Oct4-a</i> genes. Additionally, this study found that YRSSCs possess the ability to differentiate in vitro and functionally colonize in vivo within recipient bodies. This research explored the establishment of YRSSCs and their differentiation potential both in vitro and in vivo, providing a novel strategy for the genetic improvement of aquaculture fish species through germ stem cell-based gene editing and transplantation technologies.https://www.mdpi.com/2079-7737/14/5/536<i>Cyprinus carpio haematopterus</i>gonad stem cell culturespermatogenesisin vitro differentiationtransplantation
spellingShingle Huijie Zhou
Tianqi Liu
Tan Zhang
Zhipeng Sun
Huan Xu
Tingting Zhang
Yashan Yin
Na Li
Ting Yan
Youyi Kuang
Establishment and Characteristics of the Spermatogonial Stem Cell Line from the Yellow River Carp (<i>Cyprinus carpio haematopterus</i>)
Biology
<i>Cyprinus carpio haematopterus</i>
gonad stem cell culture
spermatogenesis
in vitro differentiation
transplantation
title Establishment and Characteristics of the Spermatogonial Stem Cell Line from the Yellow River Carp (<i>Cyprinus carpio haematopterus</i>)
title_full Establishment and Characteristics of the Spermatogonial Stem Cell Line from the Yellow River Carp (<i>Cyprinus carpio haematopterus</i>)
title_fullStr Establishment and Characteristics of the Spermatogonial Stem Cell Line from the Yellow River Carp (<i>Cyprinus carpio haematopterus</i>)
title_full_unstemmed Establishment and Characteristics of the Spermatogonial Stem Cell Line from the Yellow River Carp (<i>Cyprinus carpio haematopterus</i>)
title_short Establishment and Characteristics of the Spermatogonial Stem Cell Line from the Yellow River Carp (<i>Cyprinus carpio haematopterus</i>)
title_sort establishment and characteristics of the spermatogonial stem cell line from the yellow river carp i cyprinus carpio haematopterus i
topic <i>Cyprinus carpio haematopterus</i>
gonad stem cell culture
spermatogenesis
in vitro differentiation
transplantation
url https://www.mdpi.com/2079-7737/14/5/536
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