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Selection of red deer spermatozoa with different cryoresistance using density gradients

Publication at Faculty of Medicine in Pilsen |
2016

Abstract

The objective of sperm selection media is selecting the best spermatozoa and to remove seminal plasma and diluent for using them in assisted reproductive techniques (ART). It is known that individuals show different cryoresistance in response to the same freezing procedure.

Our hypothesis was that the efficacy of selection media could be dissimilar for samples with different sperm quality after thawing. Epididymal sperm samples from mature Iberian red deer were collected and frozen.

Males were classified as with high post-thaw sperm quality when sperm motility (SM) GREATER-THAN OR EQUAL TO 70%, or as with low post-thaw sperm quality when SM LESS-THAN OR EQUAL TO 69%. Samples were centrifuged using the following density gradients (DG): Percoll(R), Puresperm(R) and BovipureTM, and several functional sperm parameters were assessed after sperm selecting and washing.

Males classified with high sperm quality had higher post-thawing values (P > 0.05) for all parameters evaluated, except for linearity index (LIN), than those categorized as low sperm quality. After selection, some sperm characteristics improved (viability, apoptosis and mitochondrial activity) for both groups, showing the males with high sperm quality higher values in all sperm parameters except for kinematic traits and DNA fragmentation index (%DFI), regardless of DG.

BovipureTM yield lower values of sperm motility, viability, apoptosis and mitochondrial activity in relation to Percoll(R) and Puresperm(R) considering both quality groups. There was an interaction between the type of DG and sperm quality group for sperm viability (P = 0.040) and apoptosis (P = 0.003).

Thus, Percoll(R) selected less live and more apoptotic spermatozoa than Puresperm(R) and BovipureTM for males with low sperm quality. In conclusion, the DG are more efficient selecting spermatozoa from samples with high sperm quality, acting differently depending on initial sperm quality.