A method to obtain avian germ-line chimaeras using isolated primordial germ cells.
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 degrees 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.
Yasuda
Y
Y
Department of Anatomy, Kumamoto University Medical School, Japan.
Tajima
A
A
Fujimoto
T
T
Kuwana
T
T
D000818:Animals; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D002642:Chick Embryo; D002678:Chimera; D003370:Coturnix / Q000196:embryology; D005854:Germ Cells / Q000166:cytology / Q000637:transplantation*