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. 2021 Apr 28;100(8):101207. doi: 10.1016/j.psj.2021.101207

Figure 1.

Figure 1

General outline of producing donor-derived hatchlings with cryopreserved indigenous PGC lines. The migration of PGCs reaches its peak in the bloodstream between HH stages 13-17 (48–65 h 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 can be established. The stable cell lines need to be validated for sex, PGC specific gene expression by qPCR, PGC specific proteins by immunohistochemistry and proliferation rate by proliferation assay. For long-term storage, the cells are cryopreserved and kept in liquid nitrogen. As a next step, cryopreserved PGCs are injected into the recipient embryo. After the hatching, the presumptive germline chimaeras are backcrossed with the original breed to regenerate the donor genotype.