In vitro fertilization of bovine oocytes with semen submitted to different extenders

Revista Brasileira de Zootecnia

Coelho, Lia de Alencar¹  Almeida Junior, Ivo Luis²  Esper, César Roberto²  Garcia, Joaquim Mansano²  Vantini, Roberta² 

[1] USP;UNESP

关键词: bovine;    cleavage rate;    embryo;    in vitro fertilization;    semen extender     IntroduçãoA fecundação in vitro (FIV);    ou produção de embriões in vitro;    é uma técnica que tem sido utilizada para estudar a fisiologia dos processos de maturação ovocitária;    fecundação e desenvolvimento embrionário no estádio de pré-implantação;    bem como propiciar o desenvolvimento de outras técnicas;    como clonagem e transferência de genes;   

DOI  :  10.1590/S1516-35982000000200012

学科分类：兽医学

来源: Universidade Federal de Vicosa

【 摘 要 】

- This study investigated the effect of semen extender on in vitro development of bovine oocytes after in vitro maturation and fertilization. The ejaculate from one bull was divided in three parts which were diluted with three different extenders: Egg yolk/Lactose (EYL), Egg yolk/Citrate (EYC) and Egg yolk/Tris (EYT). Samples of semen diluted with these extenders were frozen in liquid nitrogen and, after thawing, were filtered by discontinuous Percoll gradient. The sperm concentration was adjusted to 10 x 106 cells/mL and the sperm capacitation was induced with 10 µg/mL of heparin. After 24 hours of culture for in vitro maturation, the oocytes, aspirated from ovarian follicles, were inseminated with semen diluted in TALP medium and 48 h after the zigotes were transferred into droplets prepared with TCM-199 medium containing 5% of bovine fetal serum, 5% of estrous cow serum and bovine oviduct epithelial cells in suspension under silicon oil and were further cultured of 9 days. All the cultures were performed at 38.5ºC in 5% CO2 in air. Data were analyzed by chi-square analysis. There was a significant difference in terms of cleavage rate (CR) with means of 66.0, 69.3 and 54.4% for EYL, EYC and EYT, respectively. There were no differences in terms of the morulae/blastocysts and hatching rates. These results suggest that the semen extender had no effect on in vitro development of bovine, even the CR have been affected.

【 授权许可】

CC BY-NC