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. 2019 Oct 3;9:14284. doi: 10.1038/s41598-019-50763-3

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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General outline of producing donor-derived hatchlings with sterile recipients. The migration of PGCs reaches its peak in the bloodstream between HH stages 13–17 (48–65 hours after laying in chicken); thus this is the optimal stage for collecting donor PGCs, and also this is the suitable stage for injecting them back to the recipient embryo. After the isolation, using a selective media, in vitro cultures of PGCs will be possible. For long term storage we cryopreserved the cells and keep them in liquid nitrogen. As a next step, cryopreserved or fresh PG cells are injected into the recipient embryo. After the hatching the presumptive germline chimeras are crossed back with the original breed or with each other to regenerate the donor genotype. With the usage of sterile hybrids, the treatment of the recipient embryos is not needed therefore the process is more efficient.