Animal Bioscience

  • 제38권12호
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  • Pages.2624-2634
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  • 2025
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  • 2765-0189(pISSN)
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  • 2765-0235(eISSN)
아세아태평양축산학회 (Asian Australasian Association of Animal Production Societies)
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A practical approach for the stable isolation and cultivation of chicken gonadal primordial germ cells with mitotically inactivated STO feeder cells

  • Hyeon Yang (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Bo Ram Lee (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Jae-Yeong Lee (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Keon Bong Oh (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Poongyeon Lee (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Seunghoon Lee (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Yong Jin Jo (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Haesun Lee (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Seokho Kim (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Jingu No (Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration) ;
  • Jae Yong Han (Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University) ;
  • Sung June Byun (Poultry Research Institute, National Institute of Animal Science, Rural Development Administration)
  • 투고 : 2025.03.22
  • 심사 : 2025.05.16
  • 발행 : 2025.12.01
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초록

Objective: Establishing chicken primordial germ cells (PGCs) in vitro is critical for producing genetically modified (GM) chickens. Efficient and reliable isolation and cultivation of PGCs remain significant challenges in advancing avian genetic modifications. To address these challenges, we employed a streamlined and practical approach for the efficient isolation and stable cultivation of chicken gonadal PGCs. Methods: Chicken gonadal PGCs were isolated from embryonic gonads, surgically removed and dissociated using trypsin. The PGCs were isolated by exploiting differential adhesion properties, allowing fibroblasts to attach while PGCs remained suspended. Cultivation was performed with mitotically inactivated SIM mouse embryo-derived thioguanine-resistant (STO) feeder cells under optimized culture conditions. Results: PGCs proliferated robustly, reaching over 105 cells within one month, which is comparable to previously reported methods. Characterization assays confirmed the expression of PGC-specific markers, including SSEA-1 and DAZL, along with pluripotency-related genes such as OCT4 and NANOG. Additionally, injected PGCs successfully migrated to recipient embryonic gonads, where their presence was confirmed by fluorescence analysis and PCR. Conclusion: This study highlights the effectiveness of the STO feeder-based culture system in avian germ cell research, contributing to progress in the production of germline chimeric and GM chickens.

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과제정보

This work was supported by the "Animal Science & Technology Development (Project No. PJ017202)" from the Rural Development Administration of the Republic of Korea.

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