Fertilisation of bovine oocytes by the injection of immobilised, killed spermatozoa

Department of Animal Science, Faculty of Agriculture, Kagoshima University, Japan.

Immobilised (killed) bovine spermatozoa were microinjected into bovine oocytes matured in vitro and cultured for six to nine days in vitro. A co-culture system with cumulus cells was used for the embryonic development. After one to two, three to four, five to six and seven to eight days the proportions of the oocytes which had developed to the two to four-cell, six to 12-cell, morula and blastocyst stages were 12.0 per cent (61 of 507), 9.3 per cent (47 of 507), 5.9 per cent (30 of 507) and 7.8 per cent (nine of 115), respectively. In contrast, none of the sham-operated group developed beyond the six-cell stage. This is the first report to show that bovine oocytes matured in vitro can undergo cleavage to the blastocyst stage after the injection of sperm in vitro. In addition, normal calves were obtained from the transfers of some of the embryos to recipient cows.

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