Faculty of Veterinary Medicine University of Zagreb
Abstract
In the present study we examined the effect of density gradient preparation Percoll® on bull sperm separation and in vitro fertilization (IVF) results. Frozen/thawed semen from 5 Simmental bulls was pooled and sperm quality parameters were evaluated before and after sperm separation using Percoll® density gradient preparation. We evaluated sperm motility (subjective), sperm concentration (Thoma chamber) and sperm membrane activity by HOS assay. Initial sperm parameters results were as follows: motility 50%, sperm concentration 80.80 x 106 spz/mL and functional integrity of sperm cells 39.25%. Final sperm motility was 63%, final sperm concentration 28 x 106 spz/mL and HOS assay results after Percoll® gradient was 57.60%. A total of 211 oocytes were matured and fertilized in vitro and cultured in SOFaaBSA in five repetitions. The cleavage (D2), blastocysts (D7) and hatched blastocysts (D10) were 63.13%, 20.38%, and 10.64%, respectively. We found no significant correlations between sperm parameters and IVF results (r0.05). It is possible to conclude that sperm evaluation parameters after sperm separation by gradient Percoll® are not reliable in predicting the outcome of IVF. However, using gradient Percoll® for sperm separation in IVF is necessary to improve the quality of separated sperm able to significantly improve sperm quality with a high rate of progressive motility and morphological normal spermatozoa
