A method to obtain avian germ-line chimaeras using isolated primordial germ cells

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Abstract
Summary. Primordial germ cells (PGCs), collected from the blood of 2-day-old chick embryos, were concentrated by Ficoll density centrifugation. The blood contained 0·048% PGCs and the concentrated fraction contained 3·9% PGCs in blood cells. The PGCs were picked up with a fine glass pipette, and one hundred were then injected into the terminal sinuses of 2-day-old Japanese quail embryos (24 somites); bubbles were then inserted to prevent haemorrhage. The embryos were further incubated at 38°C for 24 h, and then fixed. Serial sections were stained with the periodic acid–Schiff reagent (PAS) to demonstrate chicken PGCs and with Feulgen stain to identify quail cells. On the basis of the differences in staining properties, 63·6 ± 5·3 chick PGCs were detected in the quail embryo in the area where the gonads develop. Furthermore, 39·3 ± 4·5 chick PGCs were incorporated into the quail germinal epithelium within 24 h of the injection. A similar percentage of the host (quail) PGCs had also migrated to the germinal epithelium at the same stage of development. This technique for obtaining germ-line chimaeras will facilitate research on avian germ-line differentiation. Keywords: primordial germ cells; migration; germ-line chimaera; chick; quail; transplantation

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