Electrofusion parameters for nuclear transfer predicted using isofusion contours produced with bovine embryonic cells.
Electrofusion is a valuable technique for the nuclear transfer procedure. An enucleated oocyte is electrofused with a blastomere to create a nuclear transfer embryo. The present study constructed isofusion contours after the electrofusion of identical coupled cells that characterized all the bovine embryonic cell types used in nuclear transfer. The intersection of isofusion contours for enucleated oocytes and blastomeres provided the parameters for electrofusion during nuclear transfer. Blastomeres isolated from in vitro produced embryos 3-6 days after (in vitro fertilization) were electrofused with oocytes enucleated by centrifugation (85, 87, 89, and 73% electrofusion, respectively). The cleavage (46, 40, 37, and 28%, respectively) of the nuclear transfer embryos produced a trend that decreased as the age of the blastomeres increased. The isofusion contours provided information about the interaction between different cell types in an electric field, and gave precise electrofusion parameters for a range of bovine embryonic cell types used in nuclear transfer.
Tatham
B G
BG
Institute of Reproduction and Development, Monash University, Clayton, Victoria, Australia.
Giliam
K J
KJ
Trounson
A O
AO
D000818:Animals; D001757:Blastomeres / Q000502:physiology*; D002417:Cattle; D002459:Cell Fusion; D003584:Cytological Techniques; D053652:Nuclear Transfer Techniques