Effects of oocyte quality, semen donor and embryo co-culture system on the efficiency of blastocyst production in goats.
The aim of the study was to determine whether the selection of immature oocytes by a combination of cumulus-oocyte-complexes (COCs) morphology and staining with brilliant cresyl blue (BCB) would be helpful in selecting developmentally competent oocytes, and thereby increase the efficiency of blastocyst production from ovarian oocytes of FSH-primed, adult goats. In a second experiment the interaction between oocyte quality and semen donor was assessed. In a third experiment the usefulness of Vero cells for co-culture with goat embryos was investigated. In the pool of morphologically normal COCs recovered from ovaries following slicing (21.9+/-11.0), the mean rate of COCs classified as BCB+ was 85.6%, and the BCB- was approximately 11%. Oocytes classified as grade 1 and BCB+ exhibited the highest developmental competence (P<0.001) after in vitro maturation and fertilization compared with oocytes of grade 1 BCB- and grade 2 BCB+ or BCB-. There were no significant differences in developmental competence in grade 2 oocytes, regardless of BCB coloration. No significant differences in embryo cleavage and blastocyst formation rates among three bucks were observed when morphologically normal, BCB+ oocytes were used. For all tested bucks, differences in embryo production efficiency were related only to the oocyte quality. Similar blastocyst rates were developed from embryos co-cultured with goat oviduct epithelial cells (34.3%) and with Vero cells (33.3%). These results show that the most important criterion for selection of COCs before maturation is the visual assessment of morphological features. Staining with BCB of COCs recovered from adult goats does not enhance efficiency of selection of developmentally competent oocytes for IVF.
Katska-Ksiazkiewicz
L
L
Department of Biotechnology of Animal Reproduction, National Research Institute of Animal Production, 32-083, Balice, Poland. lkatska@izoo.krakow.pl
Opiela
J
J
Ryńska
B
B
D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D018920:Coculture Techniques; D004526:Efficiency; D046149:Embryo Culture Techniques / Q000662:veterinary*; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D006041:Goats / Q000196:embryology; D008297:Male; D009865:Oocytes / Q000166:cytology* / Q000502:physiology; D013194:Staining and Labeling; D014019:Tissue Donors; D020858:Tissue and Organ Harvesting / Q000379:methods; D014709:Vero Cells