The effect of four supplements with macromolecules on the rate of nuclear maturation, oocyte division at 48 h and embryo development at 7 days post-fertilization was evaluated. Ovaries were collected in the slaughterhouse; oocytes with two or more layers of cells were selected and matured on TCM-99 enriched with macromolecule supplements: Polyvinylpyrrolidone (PVP), polyvinylalcohol (PVA), Bovine serum albumin (BSA), Foetal bovine serum (SFB). Oocytes were cultivated at 39 °C with 5% of CO2. Nuclear maturation was evaluated at 24 hours, classifying them as germinal vesicle, metaphase I, anaphasetelophase, metaphase II and degenerated. For the evaluation of the oocyte division and embryo development, oocytes were matured under the same conditions and fertilized with spermatozoa during 18 hours in KSOM-AA culture media, and then transferred to SOF culture media for 48 h. Significant diffe...

The effect of four supplements with macromolecules on the rate of nuclear maturation, oocyte division at 48 h and embryo development at 7 days post-fertilization was evaluated. Ovaries were collected in the slaughterhouse; oocytes with two or more layers of cells were selected and matured on TCM-99 enriched with macromolecule supplements: Polyvinylpyrrolidone (PVP), polyvinylalcohol (PVA), Bovine serum albumin (BSA), Foetal bovine serum (SFB). Oocytes were cultivated at 39 °C with 5% of CO2. Nuclear maturation was evaluated at 24 hours, classifying them as germinal vesicle, metaphase I, anaphasetelophase, metaphase II and degenerated. For the evaluation of the oocyte division and embryo development, oocytes were matured under the same conditions and fertilized with spermatozoa during 18 hours in KSOM-AA culture media, and then transferred to SOF culture media for 48 h. Significant diffe...

The study was carried out to evaluate the effect of incubation time on nuclear maturation and cleavage rate of alpaca oocytes after 72 hours post-fertilization. Cumulusoocyte complexes (CCOs) were collected from ovaries collected at slaughterhouse and transported in saline solution 0.9% with antibiotic antimycotic at 35 ºC. CCOs were aspirated from 2-6 mm follicles. Experiment 1: 502 oocytes were distributed in four maturation times (30, 34, 38, 42 hours), matured in TCM-199 supplemented with 10% fetal calf serum (FCS) (v:v), 0.5 μg/mL FSH, 10 μg/mL hCG, 0.2 mM sodium pyruvate, 50 μg/mL gentamicine and 1 μg/mL oestradiol, and cultivated at 39 ºC in an atmosphere of 5% CO2 and high humidity. After maturation, CCOs were removed from maturation medium and washed with PBS supplemented with 10% FCS and 1 mg/ml of hyaluronidase, and fixed in ethanol and acetic acid (3:1). Oocytes were pl...

Se evaluó el efecto del tiempo de cultivo sobre la tasa de maduración nuclear y tasa de división post-fecundación a 72 horas de ovocitos de alpacas. Complejos Cumulus-Ovocitos (CCOs) fueron obtenidos de ovarios procedentes de animales beneficiados en el camal y transportados a 35 °C en solución salina 0.9% suplementada con antibiótico antimicótico. Los CCOs fueron aspirados de folículos de 2 a 6 mm. Experimento 1: 502 ovocitos fueron distribuidos en cuatro tiempos de maduración (30, 34, 38 y 42 horas) y madurados en TCM-199 suplementado con 10% suero fetal bovino (SFB), 0.5 µg/mL de FSH, 10 µg/mL de hCG, 0.2 mM de piruvato de sodio, 50 µg/mL de gentamicina y 1 µg/mL de E2, y cultivados a 39 ºC bajo una atmósfera de 5% de CO2 y alta humedad. Posteriormente, los ovocitos fueron removidos, lavados con PBS suplementado con 10% de SFB y 1 mg/ml de hialuronidasa y fijados en so...