Effects of selection by the Percoll density gradient method on motility, mitochondrial membrane potential and fertility in a subpopulation of Atlantic salmon (Salmo salar) testicular spermatozoa

Bravo, Williams; Lissabet, Jorge Beltrán; Jara-Seguel, Pedro; Romero, Jaime; Farías, Jorge G.; Risopatrón, Jennie; Valdebenito, Iván; Figueroa, Elías

doi:10.1016/j.anireprosci.2020.106344

Cited by 9 publications

(5 citation statements)

References 22 publications

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“…Another study conducted in sturgeon includes the evaluation of sperm motility parameters and sperm characteristics after gradient centrifugation method of control and frozen sperm samples [165]. In a freeze-thaw washing study performed with testicular sperm, the fertilization rate was found to be higher than the control [170]. In the only study in which sperm washing was used to remove bacterial load, it was observed that gradient centrifugation method reduced bacterial load, although it caused a decrease in motility rates in rainbow trout sperm [155].…”

Section: Sperm Washing and Cell Separating Methodsmentioning

confidence: 99%

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Cryopreservation Studies in Aquaculture from Past to Present: Scientific Techniques and Quality Controls for Commercial Applications

Ekici¹,

Yamaner²,

Demircan³

2023

Biomedical Engineering

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In this section, cryopreservation of fish genetic resources, which is one of the important applications to ensure the sustainability of genetic resources of freshwater fish species, is discussed. At the same time, information is provided about the possible sources of contamination that may be encountered during cryopreservation applications. In this context, the results of sperm, egg, and embryo cryopreservation studies of fish and their success and failure in applications were evaluated in addition to the process from past to present. Information is given about the contamination that may develop depending on the applications in the process of cryopreservation and dissolving processes, as well as the studies carried out to eliminate extracellular disease agents. In the section, in addition to the evaluation of the results of scientific studies, commercial companies that commercially carry out gamete cryopreservation applications are also included. The contamination that may develop depending on the applications in the process of cryopreservation and thawing processes, as well as the studies carried out to eliminate extracellular disease agents are mentioned.

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Section: Sperm Washing and Cell Separating Methodsmentioning

confidence: 99%

“…In studies using the density gradient centrifugation method, percoll was the most widely used gradient-forming chemical [169,170]. Sil-Select Plus TM [155] and AllGrad® 90% [167], which are commercial solutions produced in humans, have been tried in fish and successful results have been obtained.…”

Section: Sperm Washing and Cell Separating Methodsmentioning

confidence: 99%

Cryopreservation Studies in Aquaculture from Past to Present: Scientific Techniques and Quality Controls for Commercial Applications

Ekici¹,

Yamaner²,

Demircan³

2023

Biomedical Engineering

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“…The density gradient centrifugation technique has been used for auxiliary tool, in assisted reproduction in mammals [5] , [12] , and lastly in fish [11] for examplo, with Salmo salar [2] , allowing a proportion of sperm with greater motility, speed, and fertilization capacity. For the shaping of gradients, silica-based solutions are used, which can be coated with polyvinylpyrrolidone (PVP) [1] , or with silane [12] .…”

Section: Methods Detailsmentioning

confidence: 99%

Sperm selection of cryopreserved milt of catfish (Rhamdia quelen) by density gradient centrifugation with AllGrad® 90%

et al. 2021

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Density gradient centrifugation is a technique used to wash or separate samples of cryopreserved milt, mainly in humans and bovines allowing, for example, reducing the concentration of cryoprotectants or choosing the best portion of sperm. The proposed method seeks to reduce the presence of cryoprotectant in the cryopreserved milt of the Rhamdia qhelen and to obtain a fraction of better quality sperm. Gradient centrifugation was formed from 90% AllGrad® and different centrifugation times and forces were compared. The separated sperm presented a low increase in motility and decreased head damage and presence of gout, however, it was better compared to the non-separated samples. The speed of 1000 × g for 10 min, 4 °C, allowed 22.25 ± 4.64% of normal spermatozoa, that is, 9.25% more than the non-centrifuged milt ( p = 0.0013). The centrifugation method allows a fraction of spermatozoa morphologically less affected by cryopreservation. Density gradient centrifugation with AllGrad® 90% is proposed as a tool of easy adaptation and application for the separation of cryopreserved sperm of R. quelen . Density gradient centrifugation method at 1000 × g for 10 min allows obtaining a better fraction of normal sperm.

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“…Motility was evaluated using values from 0 to 100%, according to Magnotti et al (2022) [41]. Sperm concentration was determined with a Neubauer hemocytometer in a standard culture medium (Cortland ® ) for fish spermatozoa [42]. Samples that showed >80% motility and sperm concentrations 10 × 10 9 ± 1.4 spermatozoa/mL were included in this study [42].…”

Section: Semen Collection and Analysismentioning

confidence: 99%

Novel Apoplastic Antifreeze Proteins of Deschampsia antarctica as Enhancer of Common Cell Freezing Media for Cryobanking of Genetic Resources, a Preliminary Study

Short,

Zamorano,

Aranzaez-Ríos

et al. 2024

Biomolecules

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Antifreeze proteins (AFPs) are natural biomolecules found in cold-adapted organisms that lower the freezing point of water, allowing survival in icy conditions. These proteins have the potential to improve cryopreservation techniques by enhancing the quality of genetic material postthaw. Deschampsia antarctica, a freezing-tolerant plant, possesses AFPs and is a promising candidate for cryopreservation applications. In this study, we investigated the cryoprotective properties of AFPs from D. antarctica extracts on Atlantic salmon spermatozoa. Apoplastic extracts were used to determine ice recrystallization inhibition (IRI), thermal hysteresis (TH) activities and ice crystal morphology. Spermatozoa were cryopreserved using a standard cryoprotectant medium (C+) and three alternative media supplemented with apoplastic extracts. Flow cytometry was employed to measure plasma membrane integrity (PMI) and mitochondrial membrane potential (MMP) postthaw. Results showed that a low concentration of AFPs (0.05 mg/mL) provided significant IRI activity. Apoplastic extracts from D. antarctica demonstrated a cryoprotective effect on salmon spermatozoa, with PMI comparable to the standard medium. Moreover, samples treated with apoplastic extracts exhibited a higher percentage of cells with high MMP. These findings represent the first and preliminary report that suggests that AFPs derived from apoplastic extracts of D. antarctica have the potential to serve as cryoprotectants and could allow the development of novel freezing media.

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Effects of selection by the Percoll density gradient method on motility, mitochondrial membrane potential and fertility in a subpopulation of Atlantic salmon (Salmo salar) testicular spermatozoa

Cited by 9 publications

References 22 publications

Cryopreservation Studies in Aquaculture from Past to Present: Scientific Techniques and Quality Controls for Commercial Applications

Cryopreservation Studies in Aquaculture from Past to Present: Scientific Techniques and Quality Controls for Commercial Applications

Sperm selection of cryopreserved milt of catfish (Rhamdia quelen) by density gradient centrifugation with AllGrad® 90%

Novel Apoplastic Antifreeze Proteins of Deschampsia antarctica as Enhancer of Common Cell Freezing Media for Cryobanking of Genetic Resources, a Preliminary Study

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