Biopsy and sex determination by PCR of IVF bovine embryos

Abstract

Sex of early bovine embryos was determined by polymerase chain reaction (PCR) using a single blastomere removed at the 16-32 cell stage. Embryos were produced in vitro and biopsied on the fifth day after in vitro fertilization. Biopsied embryos were cultured on a cumulus cell monolayer until embryo transfer. For the PCR, one pair of bovine-specific and one pair of Y-chromosome-specific primers were used. Definite signals following PCR amplification were obtained in 95.4% of cases indicating that one blastomere from a preimplantation bovine embryo is sufficient for sex determination by PCR. Nineteen biopsied embryos of predetermined sex were transferred into synchronized recipient females to examine their developmental potential in vivo. Ten of the recipients (52.6%) were found to be pregnant by ultrasonography 25 days after transfer. This result did not differ significantly from that achieved with the use of the control non-manipulated IVF embryos (54.1%; P > 0.1).