Patent ID: 6600087
Filing Date: 2003-07-29
Classification: A01K,C12N

Abstract:
A process for the production of a bovine embryo comprising the following steps:(a) obtaining a bovine fetal fibroblast comprising a nucleus; (b) combining the nucleus of the fetal fibroblast with a bovine enucleated oocyte, the fetal fibroblast not being fixed by external manipulation in the GO phase before the combination, to produce a first karyoplast-cytoplast complex (KCC); (c) activating the first KCC by, (i) incubating the first KCC in medium comprising about 7% ethanol for about 5 minutes; (ii) incubating the first KCC of Step (c)(i) in a medium containing cycloheximide and an effective amount of microtubule aggregation inhibitor; (d) growing the activated KCC of step (c)(ii) in culture medium so that an embryo forms; (e) obtaining a cell from the embryo of step (d) comprising a nucleus; (f) combining the nucleus of the cell of step (e) with a bovine enucleated oocyte to produce a second KCC; (g) activating the second KCC by, (i) incubating the second KCC in medium comprising about 7% ethanol for about 5 minutes; (ii) incubating the second KCC of Step (g)(i) in a medium containing cycloheximide and an effective amount of microtubule aggregation inhibitor; (h) growing the activated KCC of step (g)(ii) in culture medium so that blastocyst form; and optionally (i) implanting the blastocyst into a female bovine thereby permitting development of a fetus from the blastocyst.