Primordial germ cell transplantation into X-ray-sterilized roosters for viable chicken production: a pilot study

Different approaches have been developed for producing viable chickens by transplanting primordial germ cells (PGCs) into recipients. Implanting PGCs into chicken embryos requires intricate techniques and results in low germline transmission efficiency. While the use of gamma-ray irradiation to produce sterilized recipients has been well established, concerns over international terrorism involving radioactive sources and facility costs have driven a shift toward X-ray irradiation in biomedical research. To date, no study has demonstrated the efficacy of X-ray irradiation in the production of sterile adult roosters for PGC transplantation. Here, we establish a practical and reliable X-ray-based sterilization method by administering 5 × 8 Gy irradiation, which effectively induces long-term spermatogenic arrest. Following PGC transplantation, spermatogenesis was restored within eight weeks, as evidenced by SSEA-1 re-expression and the production of viable offspring. Genotyping and phenotyping confirmed that all progeny were exclusively derived from donor PGCs. In summary, we demonstrate the feasibility of using X-ray irradiation to generate sterile recipient roosters for PGC transplantation. Although futher optimization is needed to improve germline transmission efficiency, this approach provides a safer, more accessible alternative to gamma-ray irradiation, with potential applications in avian biotechnology.