A comparison of BoviPure and Percoll on bull sperm separation protocols for IVF.
In the present study, we have examined the effect of density gradient preparations BoviPure and Percoll on bull sperm separation and the in vitro fertilization (IVF) and culture (IVC) results. Frozen/thawed semen from five simmental bulls were pooled. Sperm quality parameters such as sperm motility, concentration, membrane activity (HOS assay), membrane integrity (SYBR-14/PI assay) and acrosomal status (EthD-1/FITC-PSA assay) were evaluated before and after sperm processing for IVF using BoviPure and Percoll density gradient separations. The results of the evaluated parameters before sperm processing were: motility 50%, concentration 82.33x10(6)spz/mL, membrane activity 39.05%, membrane integrity 42.97% and the acrosomal status 46.90% of the live spermatozoa with intact acrosomes. After sperm processing with BoviPure and Percoll the motility was 66.67 and 64.17%, the concentration was 25.50x10(6) and 27.67x10(6)spz/mL, the membrane activity was 53.78 and 56.58%, the membrane integrity was 70.85 and 68.76% of and the acrosomal status was 74.16 and 67.46% of the live spermatozoa with intact acrosomes, respectively. Percentages were referred to the total number of spermatozoa. There were significant differences (P<0.05) between the evaluated parameters before and after sperm processing for both separation protocols. We found no significant differences (P>0.05) regarding sperm evaluation parameters between the protocols. A total of 492 oocytes were matured and fertilized in vitro and cultured in SOFaaBSA in six replicates. The cleavage (D2) and blastocysts (D7) rate were significantly higher (P<0.05) for the BoviPure group compared to the Percoll group: 75.80 and 28.21%; 61.58 and 20.83%, respectively. However, the number of hatched blastocysts (D10) did not differ significantly between sperm separation protocols (P>0.05). Our results indicate that both protocols gave suitable sperm for IVF. This finding and the similarity between those two density gradient preparations suggests that BoviPure is a good alternative for sperm separation in bovine IVF.
Samardzija
M
M
Clinic for Obstetrics and Reproduction, Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, 10000 Zagreb, Croatia. smarko@vef.hr
Karadjole
M
M
Matkovic
M
M
Cergolj
M
M
Getz
I
I
Dobranic
T
T
Tomaskovic
A
A
Petric
J
J
Surina
J
J
Grizelj
J
J
Karadjole
T
T
D000818:Animals; D002417:Cattle; D002469:Cell Separation / Q000379:methods / Q000662:veterinary*; D002499:Centrifugation, Density Gradient / Q000379:methods / Q000662:veterinary; D046149:Embryo Culture Techniques / Q000662:veterinary; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D007202:Indicators and Reagents; D008297:Male; D011205:Povidone / Q000633:toxicity; D012821:Silanes; D017640:Silicates; D012822:Silicon Dioxide / Q000633:toxicity; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology*
C509975:BoviPure; D007202:Indicators and Reagents; D012821:Silanes; D017640:Silicates; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone