EFFECT OF BOVINE SPERM CAPACITATION METHOD ON CONCENTRATION AND RATE OF in vitro FERTILIZATION
Keywords:
Cryoprotective, post mortem oocytes, maturation, motile sperm fraction, embryo.Abstract
Mammalian sperm are not able to fertilize the oocyte immediately; they must first undergo a preparation period called sperm capacitation, which is defined as the set of modifications occurring after sperm maturation in the epididymis, giving it the ability to fertilize the oocyte. A first requirement in the process of preparing bovine sperm for in vitro fertilization (IVF) includes the separation of the sperm from the diluent and/or cryoprotectant. This work was carried out at the Biotechnology Laboratory of IDIAP in order to examine the effect of sperm capacitation method on concentration and fertilization rate of in vitro fertilization. For this purpose, oocytes from post-mortem ovaries were colected and transported to the laboratory where the antral follicles (between 2 mm and 5 mm) were punctured. Subsequently, oocytes with more than 5 layers of cumulus cells were selected and matured for 20 hours. After this time, IVF was carried out. The Gradient methods of Percoll® and Swim up were used for the separation of the motil sperm fraction. Sperm concentration was measured using a SpermaCue® and the fertilization rate was evaluated at 18 hours post-fertilization using a microscope with an increase of 100x. The results indicated that the highest sperm concentration and fertilization rate were observed with Swim up (4,8 x 106 and 83,7 x 7,8 % respectively), while the values with the Percoll Gradient were 3,5 x106 and 60,22 x 6,3% for sperm concentration and fertilization rate. The total production of embryos with both methods was 95 transferable embryos, with a development rate of 21,7%. This study reported that sperm motility did not influence the fertilization rate and concludes that both sperm capacitation methods are suitable for in vitro embryo production.
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