Pre-selection by double layer density gradient centrifugation improves the fertilising capacity of frozen–thawed, capacitated stallion sperm

Morató, Roser; Soares, Juleide M. De Souza; Orero, Guifré; Mogas, T.; Miró, Jordi

doi:10.1016/j.anireprosci.2013.04.005

Cited by 11 publications

(4 citation statements)

References 48 publications

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“…Caffeine has been shown to induce hyperactivation in bovine sperm and to increase tyrosine phosphorylation and acrosomal exocytosis after induction with lysophosphatidylcholine (LPC) treatment (Marquez & Suarez, ). In stallion sperm, caffeine has been used as a sperm capacitation treatment in IVF studies with 11% (Del Campo, Donoso, Parrish, & Ginther, ) and 4.3% (Morato, Soares, Orero, Mogas, & Miro, ) of fertilized oocytes being reported and up to 49% in hamster zona pellucida‐free oocytes (Matsukawa, Takahashi, & Hanada, ). Caffeine also increased tyrosine phosphorylation levels and acrosomal exocytosis induced by LPC (Hochi, Korosue, Choi, & Oguri, ; Pommer, Rutllant, & Meyers, ).…”

Section: Discussionmentioning

confidence: 99%

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Arroyo-Salvo

Sanhueza

Fuentes

et al. 2018

Reprod Domestic Animals

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Conventional in vitro fertilization has not yet been implemented in the equine species. One of the main reasons has been the inability to develop a culture medium and incubation conditions supporting high levels of stallion sperm capacitation and hyperactivation in vitro. Although different culture media have been used for this purpose, human tubal fluid (HTF) medium, widely used in the manipulation of human and mice gametes, has not been reported so far in stallion sperm culture. The first part of this study aimed to compare HTF and Whitten's media on different stallion sperm quality and capacitation variables. Additionally, the effect of procaine, aminopyridine and caffeine in both media was evaluated on sperm motility parameters at different incubation times. Integrity and destabilization of the plasma membrane were evaluated by merocyanine 540/SYTOX Green (MC540), mitochondrial membrane potential (∆Ψm) using tetramethylrhodamine methyl ester perchlorate (TMRM), acrosome membrane integrity by PNA/FITC and tyrosine phosphorylation by P-tyrosine mouse mAb conjugated to Alexa Fluor® by flow cytometry. Motility parameters were evaluated using the integrated semen analysis system (ISAS®). We found no differences between Whitten's and HTF media and incubation time in terms of sperm viability, uninduced acrosome membrane damage or mitochondrial membrane potential at 30- and 120-min incubation. Membrane fluidity (MC540) increased in both media at 30- and 120-min incubation compared to noncapacitating conditions. Similarly, tyrosine phosphorylation increased in both media in capacitating conditions at 2- and 4-hr incubation compared to noncapacitating conditions. Although procaine showed the best result in terms of sperm hyperactivated motility in both media, aminopyridine also showed parameters consistent with the hyperactivation including an increase in curvilinear velocity and decrease in straightness. In conclusion, HTF medium and aminopyridine equally support capacitation-related parameters in stallion sperm.

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Section: Discussionmentioning

confidence: 99%

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Arroyo-Salvo

Sanhueza

Fuentes

et al. 2018

Reprod Domestic Animals

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“…These results agree with those reported by Gellert‐Mortimer, Clarke, Gordon, Hyne, and Johnston () who found a relation between the sperm motility before and after selection. However, several researchers have showed that selection techniques are more effective when sperm quality is low (Maxwell et al., ; Morató, de Souza Soares, Orero, & Mogas, ; Santiago‐Moreno et al., ), being unable to improve those sperm samples with high quality before selection (Rodriguez‐Martinez, Larsson, & Pertoft, ). It is possible that sperm samples with poor post‐thaw sperm quality are more susceptible to the selection and centrifugation procedures and for this reason never reach the values obtained from samples with high post‐thaw sperm quality using similar selection protocols.…”

Section: Discussionmentioning

confidence: 99%

Selection of red deer spermatozoa with different cryoresistance using density gradients

García-Álvarez

Soler²,

Maulen³

et al. 2016

Reprod Domestic Animals

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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. 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) ≥ 70%, or as with low post-thaw sperm quality when SM ≤ 69%. Samples were centrifuged using the following density gradients (DG): Percoll , Puresperm and Bovipure , and several functional sperm parameters were assessed after sperm selecting and washing. Males classified with high sperm quality had higher post-thawing values (p > .05) for all parameters evaluated, except for linearity index, 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. Bovipure yield lower values of sperm motility, viability, apoptosis and mitochondrial activity in relation to Percoll and Puresperm considering both quality groups. There was an interaction between the type of DG and sperm quality group for sperm viability (p = .040) and apoptosis (p = .003). Thus, Percoll selected less live and more apoptotic spermatozoa than Puresperm and Bovipure 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.

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“… 30 This technique has a beneficial role in selecting sperm cells with intact DNA and properly condensed chromatin, 31 and decreases the rates of DNA fragmentation in spermatozoa recovered from both fertile and infertile male populations 32 33 as well as in frozen-thawed spermatozoa. 34 35 Collagen and extracellular matrix components inversely affect the ICSI procedure in the final preparations, as they make the solution dense and hinder micro-manipulation. Gradient centrifugation has another advantage, eliminating compounds that could be freed during tissue grinding.…”

Section: Discussionmentioning

confidence: 99%

A laboratory modification to testicular sperm preparation technique improves spermatogenic cell yield

et al. 2014

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Testicular sperm extraction is a common procedure used to find spermatogenic cells in men with nonobstructive azoospermia. The laboratory processing of biopsied testicular tissues needs to be performed meticulously to acquire a high yield of cells. In this study, the effectiveness of mincing the tissues after testicular biopsy was assessed using histological evaluation, as was the possible adverse effect of residual tissue on the migration of spermatogenic cells during density gradient centrifugation. Our results indicate that testicular residual tissue, when laid on the density gradient medium along with the sperm wash, hinders the spermatogenic cells’ forming a pellet during centrifugation, and therefore impairs the intracytoplasmic sperm injection procedure. Whereas the mean number of recovered cells from the sperm wash medium (SWM) with residual tissue is 39.435 ± 24.849, it was notably higher (60.189 ± 28.214 cells) in the SWM without minced tissues. The remaining tissue contained no functional seminiferous tubules or spermatogenic cells in histological sections. In conclusion, the remaining residual tissue after mincing biopsied testicular tissue does not add any functional or cellular contribution to spermatogenic cell retrieval; in fact, it may block the cellular elements in the accompanying cell suspension from migrating through the gradient layers to form a pellet during centrifugation and cause loss of spermatogenic cells.

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Pre-selection by double layer density gradient centrifugation improves the fertilising capacity of frozen–thawed, capacitated stallion sperm

Cited by 11 publications

References 48 publications

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Effect of human tubal fluid medium and hyperactivation inducers on stallion sperm capacitation and hyperactivation

Selection of red deer spermatozoa with different cryoresistance using density gradients

A laboratory modification to testicular sperm preparation technique improves spermatogenic cell yield

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