Germ cell nuclei of male fetal mice can support development of chimeras to midgestation following serial transplantation.
Chimeric embryos between fertilized eggs from F1 (C57BL x CBA) and 15.5-16.5 days post coitum (dpc) male fetal germ cells (FGCs) from CD-1 strain (glucose phosphate isomerase, Gpi-1a/a) mice were produced by nuclear transfer. Briefly, a single FGC was fused with enucleated oocytes and activated, and the reconstituted oocytes were cultured to the 2-cell stage. The nucleus from the reconstituted 2-cell embryos was then transferred into an enucleated blastomere of the same stage embryos derived from F1 mice to produce chimeric embryos. The reconstituted 2-cell embryos, which synchronously divided to the 4-cell stage after treatment with nocodazole, were further cultured in vitro. Compacted morula and blastocysts were transferred to the uteri of pseudopregnant female mice. Some recipients were allowed to develop to term and the others were killed at mid gestation to analyze the contribution of donor FGC-derived cells. Survival to term was low with no chimeric animals. Glucose phosphate isomerase (GPI) analysis at midgestation revealed that some conceptuses had chimerism in the fetuses, trophoblast and yolk sac at day 10.5 of pregnancy. The contribution of donor cells was 37-47%, 19-65% and 12-63%, respectively. It was concluded that the nucleus from 15.5-16.5 dpc male fetal germ cells had the potency to develop into fetus, trophoblast and yolk sac after serial nuclear transfer with oocytes and fertilized embryos. The reason for the low viability of chimeric embryos is discussed.
Kato
Y
Y
Laboratory of Animal Reproduction, College of Agriculture, Kinki University, Nara, Japan.
Tsunoda
Y
Y
D000818:Animals; D002467:Cell Nucleus / Q000502:physiology; D002478:Cells, Cultured; D004624:Embryo Transfer; D005314:Embryonic and Fetal Development; D005260:Female; D005328:Fetal Viability; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008815:Mice, Inbred Strains; D053652:Nuclear Transfer Techniques; D013094:Spermatozoa / Q000502:physiology* / Q000648:ultrastructure; D018183:Transplantation Chimera