Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model

Abstract

To cite this article: Manuel García-Herreros & Claudia L. V. Leal (2014) Comparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model, Systems Biology in Reproductive Medicine, 60:6, 338-347, DOI: 10.3109/19396368.2014 RESEARCH ARTICLEComparative study of sperm washing and selection methods after cryopreservation and its influence on sperm subpopulational structure in a bovine model AbstractThe effect of different sperm washing-selection methods on sperm morphometric characteristics as a study to detect differences in the subpopulational structure has been carried out in detail in a bovine model. Cryopreserved sperm samples from 5 bulls were thawed, pooled, and processed by TALP-washing centrifugation method (TWCM), selective Percoll discontinuous density-gradient centrifugation method (PDGM), and self-migration swim-up separation method (SUMM). Live-dead assay (SYBR-14/ethidium homodimer-1), chlortetracycline assay (CTC), and sperm motility were assessed, and aliquots of sperm were processed for automated sperm morphometry analysis (ASMA) simultaneously before (raw thawed sperm used as control, RTS) and after different sperm washing-selection techniques. Deleterious effects of different methods were evident, particularly on sperm membrane integrity (p50.05) and capacitation status (p50.05). Moreover, each cell was measured for four primary dimensional parameters, and three shape parameters. All sperm morphometric parameters evaluated were analyzed by principal component analysis (PCA) and multivariate clustering analyses. PCA revealed two principal components for each sperm washing or separation method explaining more than the 91% of the variance. The number of subpopulations found was the same for all methods (four) except for PDGM (three). However, irrespective of the number of subpopulations defined by PCA and clustering analyses, the sperm subpopulational structure was found to be different and strongly influenced by the sperm selection procedure due to statistical differences found regarding the sperm biophysical changes induced by each method used (p50.001). It is concluded that different sperm washing-selection methods commonly used during IVF process, may lead to alterations in sperm morphometric characteristics, which might explain the different results seen after IVF, since an important influence of these methods on sperm subpopulational structure has been demonstrated.

Effect of Swim-Up and Glass Wool Techniques, With Adding Antioxidants to Tris Extender on Improving Post-Cryopreserved Some Semen Attributes of Low Semen Quality for Holstein Bulls

“…Our results were improved sperm's cell motility percentage by using swim-up and glass wool sperm separation techniques. These results were agreed with the previous results of (6,10,27,31,36,38,56) in frozen thawed cattle bull, buffalo bull, fresh ram semen, and fresh human semen. The current results were also agreed with (1) finding who found a reduction in total abnormalities percentage between swim-up filtration and control group (73.0±2.49 vs. 56.50±1.50%) of frozen thawed buffalo bull semen.…”

Sperm selection techniques in cattle: Microfilter device versus conventional methods

“…Moreover, these head size variations of spermatozoa can be affected by the species (Bos taurus vs. Bos indicus) (Yániz et al, 2016), sperm subpopulation (Valverde et al, 2016), without changing their ability to fertilize. In our study, sperm head measurements showed no significant differences ( p > 0.05) amongst sperm selection techniques, which differs from the results reported by García‐Herreros, & Leal (2014).…”

Normozoospermic versus teratozoospermic domestic cats: differential testicular volume, sperm morphometry, and subpopulation structure during epididymal maturation

“…The next step was to perform a nonhierarchical analysis using the k-means model, which uses Euclidean distances from the quantitative variables after standardization of the data, so the cluster centers were the means of the observations assigned to each cluster. The multivariate k-means cluster analysis was for classifying the spermatozoa into a reduced number of subpopulations (clusters) according to their morphometric descriptors, as described by García-Herreros and Leal 23. ANOVA and Chi-squared test procedures were applied to evaluate statistical differences in the distributions of observations (individual spermatozoa) within the donor group and subpopulations (percentages of spermatozoa assigned), and then a GLM procedure was used to determine the effects of each donor group, as well as their variation, on the relative distribution frequency of spermatozoa within the subpopulations.…”