Se utilizó plasma seminal equino en un dilutor de lactosa-EDTA para la criopreservación de espermatozoides epididimarios de equinos. Se trabajó con 12 pares de testículos de caballos beneficiados. Se separaron los epidídimos y se utilizó la técnica de lavado retrógrado para obtener los espermatozoides, inyectando 10 ml del dilutor lactosa-EDTA por el conducto deferente. Se utilizaron las muestras con más de 30% de motilidad proresiva. Las muestras fueron diluidas 1:1 con el diluyente lactosa-EDTA-plasma seminal y se envasó en pajillas de 0.5 ml a una concentración 386.3 x 106, y fueron congeladas en nitrógeno líquido y almacenadas por 10 días. Los valores obtenidos para las muestras frescas y descongeladas fueron: motilidad progresiva: 43.3 y 16.4% (p<0.05), viabilidad: 48.3 y 40.5%, morfología normal: 67.1 y 56.5%, e integridad de la membrana plasmática (HOS): 48.3 y 45.5%
Equine seminal plasma was used in a lactose-EDTA extender for cryopreservation of equine epididymal sperm. Twelve pairs of testicles of slaughtered horses were used. Epididymides were separated and washed applying the retrograde technique to obtain sperm by injecting 10 ml of lactose-EDTA extender through the vas deferens. Samples with more than 30% motility were used. Samples were diluted 1: 1 with the lactose-EDTAxtender seminal plasma and filled into 0.5 ml straws at a concentration of 386.3 x 106, frozen in liquid nitrogen and stored for 10 days. The values for fresh and thawed samples were: motility: 43.3 and 16.4% (p<0.05), viability: 48.3 and 40.5%, normal morphology: 67.1 and 56.5%, and integrity of the plasma membrane (HOS): 48.3 and 45.5%
The aim of in vitro maturation oocyte systems is to produce oocytes of comparable quality to those derived in vivo. The present study was designed to examine the surface morphological changes of the cumulus-oocyte complex (COC) and nuclear maturation in a culture system containing pyruvate. Ovaries were obtained from a slaughterhouseand transported to the laboratory within 2 h at 35-39ºC,and rinsed three times in 0.9% NaCl. The COCs were harvested from the ovaries and in vitro maturation was evaluated in San Marcos (SM) medium, a chemically defined culture system containing 22.3 mM sodium pyruvate. Oocytes were cultured in SM, SM + porcine follicular fluid (pFF) and in SM + pFF + gonadotropins (eCG and hCG) for 20-22 h and then without hormonal supplements for an additional 20-22 h. After culture, the degree of cumulus expansion and frequency of nuclear maturation were determined. Oocytes matured in SM (40.9%) and SM + pFF (42.9%) showed moderate cumulus expansion, whereas oocytes matured in SM + pFF + gonadotropins (54.6%) showed high cumulus expansion. The maturation rate of cultured oocytes, measured in function of the presence of the polar corpuscle, did not differ significantly between SM (40.9 ± 3.6%) and SM + pFF (42.9 ± 3.7%). These results indicate that pig oocytes can be successfully matured in a chemically definedmedium and suggest a possible bifunctional role of pyruvate as an energy substrate and as an antioxidant protecting oocytes against the stress of the in vitro environment.