[In vitro fertilization of bovine and sheep follicles oocytes by goat sperm]
1993
Catalan V, Alfredo Francisco
The objectives of this study were: (1) to verify in vitro fertilization in sheep and cattle oocytes with goat sperm, and (2) to study the effect of cumulus cells on the efficiency of the in vitro fertilization of sheep and cattle oocyte by goat spermatozoa. Ovaries were obtained at slaughtered house, and then ovarian follicles were aspirated and matured in culture medium HEPES 199 + 20% v/v goat oestrus serum in microdrops under mineral oil. The oocytes were maintained at 39 deg C, in 5% CO2 with humidified air for 24 hours. Fresh goat semen was obtained with artificial vagina and incubated at 20 deg C for 4h. Then sperm was washed twice with TALP medium + goat oestrus serum and then capacitated with heparin. In the first experiment, cattle and sheep matured oocytes were incubated in goat sperm preparations. In the second one, cattle and sheep oocytes enclosed or denuded from granulosa cells were incubated in sperm suspensions. After 18 hours of gamete interaction, oocytes were cleaned from cells and then stained with aceto-lacmoid to evaluate fertilization rate. The oocytes were considered fertilized when both pronuclei and sperm tail were present in the cytoplasm. The results showed that heterologous fertilization rate is high, and similar for both species (Homologous: 91.3%, cattle: 82.72%, sheep: 79.76%). In addition cumulus cells did not modify the efficiency of heterologous fertilization (Goat: 96.55 and 100% in enclosed and denuded oocytes; cattle: 74% and 88.89% and sheep: 68.42% and 78.85% respectively)
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