Analysis of selected sperm by density gradient centrifugation might aid in the estimation of in vivo fertility of thawed ram spermatozoa

García-Álvarez, Olga; Maroto-Morales, Alejandro; Ramón, Manuel; Olmo, Enrique del; Montoro, Vidal; Domínguez-Rebolledo, Álvaro E.; Bisbal, A.; Jiménez-Rabadán, Pilar; Pérez-Guzmán, M. D.; Soler, Ana Josefa

doi:10.1016/j.theriogenology.2010.04.027

Cited by 45 publications

(27 citation statements)

References 59 publications

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“…In dog semen [42], frozen-thawed Androcoll-C treated samples were enriched in subpopulations with highly active and PM compared with the initial samples. Percoll and PureSperm gradients have been used in the selection of sperm in several species to improve the quality of seminal samples (ram [43], marmoset [32], stallion [25,34], bull [31,44], dog [45], and human [29,46]). Regarding brown bear sperm, the spermatozoa selected by PureSperm gradient [28] showed a higher TM compared with the fresh sample immediately after centrifugation.…”

Section: Discussionmentioning

confidence: 99%

Effect of colloid (Androcoll-Bear, Percoll, and PureSperm) selection on the freezability of brown bear (Ursus arctos) sperm

Álvarez‐Rodríguez

Álvarez

Anel-López³

et al. 2016

Theriogenology

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Section: Discussionmentioning

confidence: 99%

Effect of colloid (Androcoll-Bear, Percoll, and PureSperm) selection on the freezability of brown bear (Ursus arctos) sperm

Álvarez‐Rodríguez

Álvarez

Anel-López³

et al. 2016

Theriogenology

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“…Then, a sperm selection by discontinuous density gradient on Percoll (1 mL of 45% Percoll over 1 mL of 90% Percoll) was carried out at room temperature (20 C-22 C). Thawed semen was layered on top of the two Percoll layers and centrifuged at 700 Â g for 10 minutes [36][37][38]. After centrifugation, the supernatant was carefully discarded.…”

Section: Experimental Designmentioning

confidence: 99%

Effect of different media additives on capacitation of frozen–thawed ram spermatozoa as a potential replacement for estrous sheep serum

García-Álvarez¹,

Maroto-Morales²,

Jiménez-Rabadán

et al. 2015

Theriogenology

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Capacitation is a key process through which spermatozoa acquire their fertilizing ability. This event is required for the successful application of assisted reproductive technologies such as IVF. The aim of the present study was to investigate the effect of using a synthetic oviductal fluid medium supplemented with either heparin-hypotaurine alone, in combination with progesterone (P4), 17β-estradiol (E2), or BSA, or just β-cyclodextrin, in replacement for estrous sheep serum (ESS) for ram sperm capacitation. After incubation in the corresponding media for 15 (time 0) or 60 minutes, sperm function was evaluated by computerized sperm motility analysis and flow cytometry (plasma membrane status and fluidity). Treatments rendering the best results in regards to sperm function parameters related to capacitation were used for an IVF test. Herein, neither heparin-hypotaurine (alone), or in combination with P4, or E2, nor β-cyclodextrin induced capacitation-related changes in frozen-thawed ram spermatozoa. Only the medium supplemented with heparin-hypotaurine-BSA was able to induce changes compatible with in vitro capacitation relating to sperm motility pattern and plasma membrane fluidity, comparable to those in ESS-containing medium. Both media yielded sperm parameter values that differed (P < 0.05) from those obtained in the rest of the media tested. However, after the IVF trial, BSA was unable to support cleavage rates (21.80%) comparable to those obtained with ESS (52.60%; P < 0.05). We conclude that heparin-hypotaurine, P4, E2, β-cyclodextrin, or BSA is not suitable for replacing ESS in capacitation and fertilization media for ram spermatozoa.

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“…A recent publication demonstrated that there is a close relationship between nuclear vacuoles and clinical outcomes after ICSI, confirming that the nuclear morphology could be considered a morphological marker of developmental competence, but without any relationship with the DNA fragmentation [46,47]. To date, does not exit any tools to select sperm with chromatin integrity.…”

Section: The Chromatin Integrity As Biological Marker Of Sperm Competmentioning

confidence: 99%

New molecular markers for the evaluation of gamete quality

Ruvolo

Fattouh

Bosco

et al. 2013

J Assist Reprod Genet

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Purpose Only 30 % of IVF cycles result in a pregnancy, so that multiple embryos need to be replaced, per treatment cycle, to increase pregnancy rates, resulting in a multiple gestation rate of 25 %. The use of new markers in the gamete selection, could reduce the number of the oocytes to be fertilized and embryos to be produced, but the tools to evidence the gamete competence remain unavailable and more studies are needed to identify bio-markers to select the best oocyte and sperm to produce embryos with higher implantation potentiality. Methods To define oocyte competence, the apoptosis of the surrounding cumulus cells and the oxygen consumption rates for individual oocytes before fertilization seems to provide a non-invasive marker of oocyte competence and hence a quantitative assessment of the reproductive potential for the oocyte. The chromatin integrity seems to be used also as biological marker of sperm competence, together with the morphological evaluation of large vacuoles in the head.Results The apoptosis rate of cumulus cells lower than 25 % and an higher oxygen consumption could be an evidence of an overall metabolic activity, related to a better fertilization ability and embryo cleavage quality. The apoptosis rate of the sperm chromatin, evaluated by direct Tunel in situ analysis, seems to be, also for the male gamete, a marker of competence and implantation potentiality, in particular when it is lower than 20 %. The evaluation of the presence of large vacuoles in the sperm head prior to perform ICSI seems to increase the implantation rate, but it is not associated to chromatin integrity. Conclusions The biological concept of competence appears unrelated to any morphological parameters, so that it is necessary to investigate new molecular markers in the gamete selection. Apoptosis of cumulus cells in the oocytes and spermatozoa, revealing the presence of large vacuoles, could help to determine the competence of the gamete to be fertilize.

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Analysis of selected sperm by density gradient centrifugation might aid in the estimation of in vivo fertility of thawed ram spermatozoa

Cited by 45 publications

References 59 publications

Effect of colloid (Androcoll-Bear, Percoll, and PureSperm) selection on the freezability of brown bear (Ursus arctos) sperm

Effect of colloid (Androcoll-Bear, Percoll, and PureSperm) selection on the freezability of brown bear (Ursus arctos) sperm

Effect of different media additives on capacitation of frozen–thawed ram spermatozoa as a potential replacement for estrous sheep serum

New molecular markers for the evaluation of gamete quality

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