Effect of different mini‐volume colloid centrifugation configurations on flow cytometrically sorted sperm recovery efficiency and quality using a computer‐assisted semen analyzer
Straws of sex‐sorted sperm are usually packaged at a low concentration (e.g., ~2.1 × 106 sperm/ml) and cost significantly more than unsorted conventional semen from the same sire. In order to maximize the efficiency of using sex‐sorted sperm under in vitro fertilization conditions, the selection of...
| Autores principales: | , , , , , |
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| Formato: | info:ar-repo/semantics/artículo |
| Lenguaje: | Inglés |
| Publicado: |
Wiley
2018
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/2879 https://onlinelibrary.wiley.com/doi/abs/10.1111/rda.13048 https://doi.org/10.1111/rda.13048 |
| _version_ | 1855035043346382848 |
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| author | Ferre, Luis Bernardo Chitwood, James L. Fresno, Cristóbal Ortega, Hugo Hector Kjelland, Michael E. Ross, Pablo J. |
| author_browse | Chitwood, James L. Ferre, Luis Bernardo Fresno, Cristóbal Kjelland, Michael E. Ortega, Hugo Hector Ross, Pablo J. |
| author_facet | Ferre, Luis Bernardo Chitwood, James L. Fresno, Cristóbal Ortega, Hugo Hector Kjelland, Michael E. Ross, Pablo J. |
| author_sort | Ferre, Luis Bernardo |
| collection | INTA Digital |
| description | Straws of sex‐sorted sperm are usually packaged at a low concentration (e.g., ~2.1 × 106 sperm/ml) and cost significantly more than unsorted conventional semen from the same sire. In order to maximize the efficiency of using sex‐sorted sperm under in vitro fertilization conditions, the selection of an appropriate sperm separation technique is essential. In this study, the effect of using different silane‐coated silica colloid dilutions and layering configurations during centrifugation of sex‐sorted sperm was examined over an extended period of incubation time. Sperm recovery and viability after centrifugation using the colloid separation technique were measured along with several sperm motility parameters using CASA. For this purpose, frozen and thawed sex‐sorted sperm samples were centrifuged using mini‐volume single‐layer (40%, 60% and 80%) and mini‐volume two‐layer (45%/90%, 40%/80% and 30%/60%) separation configurations using PureSperm®. A single layer of 40% PureSperm® recovered significantly more sex‐sorted sperm (78.07% ± 2.28%) followed by a single layer of 80% PureSperm® (68.43% ± 2.33%). The lowest sperm recovery was obtained using a two‐layer PureSperm® dilution of 45%/90% (47.57% ± 2.33%). Single‐layer centrifugation recovered more sorted sperm (68.67% ± 1.74%) than two layer (53.74% ± 1.74%) (p < .0001). A single layer of 80% PureSperm® exhibited the highest sorted sperm viability (72.01% ± 2.90%) after centrifugation (p < .05). The mini‐volume single layer of 80% PureSperm® was determined to be an effective alternative to a two‐layer centrifugation configuration for sex‐sorted sperm selection. In addition, single‐layer colloid dilution of 80% performed either as well as or significantly outperformed the other treatments, as well as the control, with regard to motility (MOT) for all time periods of analysis. |
| format | info:ar-repo/semantics/artículo |
| id | INTA2879 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| publisher | Wiley |
| publisherStr | Wiley |
| record_format | dspace |
| spelling | INTA28792022-12-20T10:50:58Z Effect of different mini‐volume colloid centrifugation configurations on flow cytometrically sorted sperm recovery efficiency and quality using a computer‐assisted semen analyzer Ferre, Luis Bernardo Chitwood, James L. Fresno, Cristóbal Ortega, Hugo Hector Kjelland, Michael E. Ross, Pablo J. Reproducción Espermatozoo Fecundación in Vitro Centrifugación Coloides Reproduction Spermatozoa In Vitro Fertilization Centrifuging Colloids Esperma Straws of sex‐sorted sperm are usually packaged at a low concentration (e.g., ~2.1 × 106 sperm/ml) and cost significantly more than unsorted conventional semen from the same sire. In order to maximize the efficiency of using sex‐sorted sperm under in vitro fertilization conditions, the selection of an appropriate sperm separation technique is essential. In this study, the effect of using different silane‐coated silica colloid dilutions and layering configurations during centrifugation of sex‐sorted sperm was examined over an extended period of incubation time. Sperm recovery and viability after centrifugation using the colloid separation technique were measured along with several sperm motility parameters using CASA. For this purpose, frozen and thawed sex‐sorted sperm samples were centrifuged using mini‐volume single‐layer (40%, 60% and 80%) and mini‐volume two‐layer (45%/90%, 40%/80% and 30%/60%) separation configurations using PureSperm®. A single layer of 40% PureSperm® recovered significantly more sex‐sorted sperm (78.07% ± 2.28%) followed by a single layer of 80% PureSperm® (68.43% ± 2.33%). The lowest sperm recovery was obtained using a two‐layer PureSperm® dilution of 45%/90% (47.57% ± 2.33%). Single‐layer centrifugation recovered more sorted sperm (68.67% ± 1.74%) than two layer (53.74% ± 1.74%) (p < .0001). A single layer of 80% PureSperm® exhibited the highest sorted sperm viability (72.01% ± 2.90%) after centrifugation (p < .05). The mini‐volume single layer of 80% PureSperm® was determined to be an effective alternative to a two‐layer centrifugation configuration for sex‐sorted sperm selection. In addition, single‐layer colloid dilution of 80% performed either as well as or significantly outperformed the other treatments, as well as the control, with regard to motility (MOT) for all time periods of analysis. EEA Rafaela Fil: Ferre, Luis Bernardo. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela; Argentina. Fil: Chitwood, James L. University of California Davis. Department of Animal Science; Estados Unidos Fil: Fresno, C. National Institute of Genomic Medicine (INMEGEN). Computational Genomics Division; México Fil: Ortega, Hugo Hector. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Laboratorio de Biología Celular y Molecular Aplicada; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Ciencias Veterinarias del Litoral. Universidad Nacional del Litoral. Facultad de Ciencias Veterinarias. Instituto de Ciencias Veterinarias del Litoral; Argentina. Fil: Kjelland, M.E. Conservation, Genetics and Biotech, LLCVicksburg; Estados Unidos Fil: Ross, Pablo J. University of California Davis. Department of Animal Science; Estados Unidos 2018-07-25T17:39:23Z 2018-07-25T17:39:23Z 2018-02 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/2879 https://onlinelibrary.wiley.com/doi/abs/10.1111/rda.13048 0936-6768 1439-0531 https://doi.org/10.1111/rda.13048 eng info:eu-repograntAgreement/INTA/PNSA/1115053/AR. Balcarce, Buenos Aires/Biotecnologías reproductivas y desarrollo de metodologías de diagnóstico, control y prevención de las enfermedades infecciosas y parasitarias que afectan la concepción, gestación y período neonatal en especies de interés zootécnico. info:eu-repo/semantics/restrictedAccess application/pdf Wiley Reproduction in Domestic Animals 53 (1) : 26-33 (February 2018) |
| spellingShingle | Reproducción Espermatozoo Fecundación in Vitro Centrifugación Coloides Reproduction Spermatozoa In Vitro Fertilization Centrifuging Colloids Esperma Ferre, Luis Bernardo Chitwood, James L. Fresno, Cristóbal Ortega, Hugo Hector Kjelland, Michael E. Ross, Pablo J. Effect of different mini‐volume colloid centrifugation configurations on flow cytometrically sorted sperm recovery efficiency and quality using a computer‐assisted semen analyzer |
| title | Effect of different mini‐volume colloid centrifugation configurations on flow cytometrically sorted sperm recovery efficiency and quality using a computer‐assisted semen analyzer |
| title_full | Effect of different mini‐volume colloid centrifugation configurations on flow cytometrically sorted sperm recovery efficiency and quality using a computer‐assisted semen analyzer |
| title_fullStr | Effect of different mini‐volume colloid centrifugation configurations on flow cytometrically sorted sperm recovery efficiency and quality using a computer‐assisted semen analyzer |
| title_full_unstemmed | Effect of different mini‐volume colloid centrifugation configurations on flow cytometrically sorted sperm recovery efficiency and quality using a computer‐assisted semen analyzer |
| title_short | Effect of different mini‐volume colloid centrifugation configurations on flow cytometrically sorted sperm recovery efficiency and quality using a computer‐assisted semen analyzer |
| title_sort | effect of different mini volume colloid centrifugation configurations on flow cytometrically sorted sperm recovery efficiency and quality using a computer assisted semen analyzer |
| topic | Reproducción Espermatozoo Fecundación in Vitro Centrifugación Coloides Reproduction Spermatozoa In Vitro Fertilization Centrifuging Colloids Esperma |
| url | http://hdl.handle.net/20.500.12123/2879 https://onlinelibrary.wiley.com/doi/abs/10.1111/rda.13048 https://doi.org/10.1111/rda.13048 |
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