Migration of primordial germ cells isolated from embryonic blood into the gonads after transfer to stage X blastoderms and detection of germline chimaerism by PCR.
1. The present study was carried out to determine whether primordial germ cells isolated from embryonic blood can enter the bloodstream and successfully migrate to the germinal ridges of recipient embryos after transfer to stage X blastoderms, and also whether they can differentiate into blood cells, as is suggested in mice. 2. Primordial germ cells were transfected in vitro by lipofection and then transferred to stage X blastoderms. The introduced GFP gene was efficiently expressed in the gonads of 6-d incubated embryos. 3. Freshly collected primordial germ cells were transferred to stage X blastoderms. The fate of the transferred primordial germ cells was traced by detecting the single nucleotide polymorphism in the D-loop region of the mitochondrial DNA in White Leghorn and Barred Plymouth Rock chickens used in this study. The transferred donor primordial germ cell-derived cells were detected in the gonads, but not in the blood cells, of 17-d incubated embryos by PCR. 4. This procedure for primordial germ cell manipulation could provide a novel method of producing germline chimaeric chickens. 5. In conclusion, our findings indicate that primordial germ cells isolated from embryonic blood can migrate to the germinal ridges of recipient embryos after being transferred to stage X blastoderms. Although these transferred primordial germ cells differentiated into germ cells, no differentiation into blood cells was observed.
Naito
M
M
Animal Genetic Engineering Laboratory, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan. mnaito@affrc.go.jp
Sano
A
A
Harumi
T
T
Matsubara
Y
Y
Kuwana
T
T
D000818:Animals; D001756:Blastoderm / Q000166:cytology*; D002454:Cell Differentiation; D002465:Cell Movement; D017690:Cell Transplantation; D002642:Chick Embryo / Q000166:cytology* / Q000254:growth & development; D005854:Germ Cells / Q000502:physiology*; D006066:Gonads / Q000196:embryology*; D049452:Green Fluorescent Proteins; D016133:Polymerase Chain Reaction; D013234:Stem Cells / Q000502:physiology*; D018183:Transplantation Chimera
D049452:Green Fluorescent Proteins