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However, most methods for this purpose in birds face serious limitations such as partial germ cell depletion, high toxicity and mortality, or the need to use expensive technologies. Here, we validated a simple and practical technique of transferring quail testicular cells into chicken testes depleted of endogenous spermatozoa by fractioned chemotherapy (20\u00a0mg\/kg\/week busulfan for 5\u00a0weeks). This protocol resulted in a very low mortality of the treated day-old chicks and, despite maintenance of androgenic activity, sperm production\u00a0was decreased by 84.3% at 25\u00a0weeks of age. NANOG immunostaining revealed that very few to no germ cells were present following treatment with 20 and 40\u00a0mg\/kg, respectively. RT-qPCR data also showed that <jats:italic>c-MYC<\/jats:italic> and <jats:italic>NANOG<\/jats:italic> expression declined in these treatments, but <jats:italic>GRF\u03b11<\/jats:italic> and <jats:italic>BID<\/jats:italic> expressions remained unaltered among groups. After xenotransplantation, quail germ cells were immunodetected in chicken testes using a species-specific antibody (QCPN), and quail ovalbumin DNA was found in seminal samples collected from chicken recipients. Together, these data confirm that fractionated administration of busulfan in hatchlings is a practical, effective, and safe protocol to prepare recipient male birds capable of supporting xenogeneic spermatogenesis.<\/jats:p>","DOI":"10.1038\/s41598-023-45019-0","type":"journal-article","created":{"date-parts":[[2024,2,7]],"date-time":"2024-02-07T03:02:00Z","timestamp":1707274920000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Successful xenotransplantation of testicular cells following fractionated chemotherapy of recipient birds"],"prefix":"10.1038","volume":"14","author":[{"given":"Marcel Henrique","family":"Blank","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Allison Jun Taguchi","family":"Kawaoku","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruno Rog\u00e9rio","family":"Rui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Claudia Oliveira","family":"Carreira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thais Rose dos Santos","family":"Hamilton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcelo Demarchi","family":"Goissis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ricardo Jos\u00e9 Garcia","family":"Pereira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,2,7]]},"reference":[{"key":"45019_CR1","doi-asserted-by":"publisher","first-page":"643","DOI":"10.1080\/00071660310001616246","volume":"44","author":"P Trefil","year":"2003","unstructured":"Trefil, P. et al. 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