Elimination of apoptotic boar spermatozoa using magnetic activated cell sorting
One of the features of apoptosis is the externalization of phosphatidylserine which could be used to remove apoptotic cells from semen preparations. Magnetic-activated cell sorting using annexin V-conjugated microbeads which bind to phosphatidylserine could be used to enhance semen quality. Twelve b...
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| Veröffentlicht in: | Acta veterinaria Brno 2014-01, Vol.83 (1), p.13-18 |
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| creator | Mrkun, J., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses Dolensek, T., University of Ljubljana (Slovenia). Veterinary Faculty Knific, T., University of Ljubljana (Slovenia). Veterinary Faculty Pislar, A., University of Ljubljana (Slovenia). Faculty of Pharmacy Kosec, M., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses Kos, J., University of Ljubljana (Slovenia). Faculty of Pharmacy Zrimsek, P., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses |
| description | One of the features of apoptosis is the externalization of phosphatidylserine which could be used to remove apoptotic cells from semen preparations. Magnetic-activated cell sorting using annexin V-conjugated microbeads which bind to phosphatidylserine could be used to enhance semen quality. Twelve boar semen samples after 3 days of liquid storage at 16ÂÂÂ17 degC were subjected to magnetic-activated cell sorting. Bound and unbound fractions and control samples were subjected to flow cytometry following the staining of spermatozoa with Annexin V conjugated with Alexa Fluor 488 and propidium iodide. Four subpopulations were obtained: live, early apoptotic live, late apoptotic, early necrotic dead and late necrotic dead. The frequency of early apoptotic and late necrotic spermatozoa was significantly higher (P less than 0.05) in bound (14.1 +/- 10.6% and 24.1 +/- 10.2%, respectively) than in unbound fractions (3.4 +/- 2.1% and 12.7 +/- 3.1%) and control (3.5 +/-+/- 1.6% and 12.0 +/- 5.0%). The lowest concentration of live spermatozoa was found in the bound fraction (10.6 +/- 8.0 %), which differed significantly (P less than 0.05) from the control. In unbound fractions there was a significantly higher concentration (P less than 0.05) of morphologically normal spermatozoa (31.8 +/- 12.6%) compared to bound ones (5.9 +/- 7.3%). A significantly (P less than 0.05) lower proportion of morphologically normal spermatozoa was observed in both fractions compared to control (67.2 +/- 17.0%). Boar spermatozoa were separated by the above method for the first time, however, the results showed this method to be inappropriate for boar semen separation under the tested conditions. |
| doi_str_mv | 10.2754/avb201483010013 |
| format | Article |
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Clinic for Reproduction and Horses ; Dolensek, T., University of Ljubljana (Slovenia). Veterinary Faculty ; Knific, T., University of Ljubljana (Slovenia). Veterinary Faculty ; Pislar, A., University of Ljubljana (Slovenia). Faculty of Pharmacy ; Kosec, M., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses ; Kos, J., University of Ljubljana (Slovenia). Faculty of Pharmacy ; Zrimsek, P., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creator><creatorcontrib>Mrkun, J., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses ; Dolensek, T., University of Ljubljana (Slovenia). Veterinary Faculty ; Knific, T., University of Ljubljana (Slovenia). Veterinary Faculty ; Pislar, A., University of Ljubljana (Slovenia). Faculty of Pharmacy ; Kosec, M., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses ; Kos, J., University of Ljubljana (Slovenia). Faculty of Pharmacy ; Zrimsek, P., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creatorcontrib><description>One of the features of apoptosis is the externalization of phosphatidylserine which could be used to remove apoptotic cells from semen preparations. Magnetic-activated cell sorting using annexin V-conjugated microbeads which bind to phosphatidylserine could be used to enhance semen quality. Twelve boar semen samples after 3 days of liquid storage at 16ÂÂÂ17 degC were subjected to magnetic-activated cell sorting. Bound and unbound fractions and control samples were subjected to flow cytometry following the staining of spermatozoa with Annexin V conjugated with Alexa Fluor 488 and propidium iodide. Four subpopulations were obtained: live, early apoptotic live, late apoptotic, early necrotic dead and late necrotic dead. The frequency of early apoptotic and late necrotic spermatozoa was significantly higher (P less than 0.05) in bound (14.1 +/- 10.6% and 24.1 +/- 10.2%, respectively) than in unbound fractions (3.4 +/- 2.1% and 12.7 +/- 3.1%) and control (3.5 +/-+/- 1.6% and 12.0 +/- 5.0%). The lowest concentration of live spermatozoa was found in the bound fraction (10.6 +/- 8.0 %), which differed significantly (P less than 0.05) from the control. In unbound fractions there was a significantly higher concentration (P less than 0.05) of morphologically normal spermatozoa (31.8 +/- 12.6%) compared to bound ones (5.9 +/- 7.3%). A significantly (P less than 0.05) lower proportion of morphologically normal spermatozoa was observed in both fractions compared to control (67.2 +/- 17.0%). Boar spermatozoa were separated by the above method for the first time, however, the results showed this method to be inappropriate for boar semen separation under the tested conditions.</description><identifier>ISSN: 0001-7213</identifier><identifier>EISSN: 1801-7576</identifier><identifier>DOI: 10.2754/avb201483010013</identifier><language>eng</language><subject>ALMACENAMIENTO EN FRIO ; ANIMALES JOVENES ; APOPTOSE ; APOPTOSIS ; BIOLOGICAL PRESERVATION ; BOARS ; BREEDS (ANIMALS) ; CELL MEMBRANES ; CELLS ; CELLULE ; CELULAS ; COLD STORAGE ; CONGELACION ; CONGELATION ; CONSERVACION BIOLOGICA ; CONSERVATION BIOLOGIQUE ; CRIOPROTECTORES ; CRYOPROTECTANTS ; CRYOPROTECTEUR ; DAMAGE ; DANOS ; DEGAT ; DISENO EXPERIMENTAL ; DISPOSITIF EXPERIMENTAL ; ESPERMATOZOO ; EVALUACION ; EVALUACION DEL IMPACTO ; EVALUATION ; EVALUATION DE L'IMPACT ; EXPERIMENTAL DESIGN ; FERTILIDAD ; FERTILITE ; FERTILITY ; FREEZING ; IMPACT ASSESSMENT ; JEUNE ANIMAL ; MEMBRANAS CELULARES ; MEMBRANE CELLULAIRE ; MOUVEMENT ; MOVEMENT ; MOVIMIENTO ; PERMEABILIDAD ; PERMEABILITE ; PERMEABILITY ; RACE (ANIMAL) ; RAZAS (ANIMALES) ; REPRODUCCION SEXUAL ; REPRODUCTION SEXUEE ; SEXUAL REPRODUCTION ; SPERMATOZOA ; SPERMATOZOIDE ; STOCKAGE AU FROID ; TEMPS ; TIEMPO ; TIME ; VERRACO ; VERRAT ; VIABILIDAD ; VIABILITE ; VIABILITY ; YOUNG ANIMALS</subject><ispartof>Acta veterinaria Brno, 2014-01, Vol.83 (1), p.13-18</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c304t-d542dfcfe8987f2b12cef07c98c59cf03035402faeb00cbbbd15770e872adc383</citedby><cites>FETCH-LOGICAL-c304t-d542dfcfe8987f2b12cef07c98c59cf03035402faeb00cbbbd15770e872adc383</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,782,786,27931,27932</link.rule.ids></links><search><creatorcontrib>Mrkun, J., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creatorcontrib><creatorcontrib>Dolensek, T., University of Ljubljana (Slovenia). Veterinary Faculty</creatorcontrib><creatorcontrib>Knific, T., University of Ljubljana (Slovenia). Veterinary Faculty</creatorcontrib><creatorcontrib>Pislar, A., University of Ljubljana (Slovenia). Faculty of Pharmacy</creatorcontrib><creatorcontrib>Kosec, M., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creatorcontrib><creatorcontrib>Kos, J., University of Ljubljana (Slovenia). Faculty of Pharmacy</creatorcontrib><creatorcontrib>Zrimsek, P., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creatorcontrib><title>Elimination of apoptotic boar spermatozoa using magnetic activated cell sorting</title><title>Acta veterinaria Brno</title><description>One of the features of apoptosis is the externalization of phosphatidylserine which could be used to remove apoptotic cells from semen preparations. Magnetic-activated cell sorting using annexin V-conjugated microbeads which bind to phosphatidylserine could be used to enhance semen quality. Twelve boar semen samples after 3 days of liquid storage at 16ÂÂÂ17 degC were subjected to magnetic-activated cell sorting. Bound and unbound fractions and control samples were subjected to flow cytometry following the staining of spermatozoa with Annexin V conjugated with Alexa Fluor 488 and propidium iodide. Four subpopulations were obtained: live, early apoptotic live, late apoptotic, early necrotic dead and late necrotic dead. The frequency of early apoptotic and late necrotic spermatozoa was significantly higher (P less than 0.05) in bound (14.1 +/- 10.6% and 24.1 +/- 10.2%, respectively) than in unbound fractions (3.4 +/- 2.1% and 12.7 +/- 3.1%) and control (3.5 +/-+/- 1.6% and 12.0 +/- 5.0%). The lowest concentration of live spermatozoa was found in the bound fraction (10.6 +/- 8.0 %), which differed significantly (P less than 0.05) from the control. In unbound fractions there was a significantly higher concentration (P less than 0.05) of morphologically normal spermatozoa (31.8 +/- 12.6%) compared to bound ones (5.9 +/- 7.3%). A significantly (P less than 0.05) lower proportion of morphologically normal spermatozoa was observed in both fractions compared to control (67.2 +/- 17.0%). Boar spermatozoa were separated by the above method for the first time, however, the results showed this method to be inappropriate for boar semen separation under the tested conditions.</description><subject>ALMACENAMIENTO EN FRIO</subject><subject>ANIMALES JOVENES</subject><subject>APOPTOSE</subject><subject>APOPTOSIS</subject><subject>BIOLOGICAL PRESERVATION</subject><subject>BOARS</subject><subject>BREEDS (ANIMALS)</subject><subject>CELL MEMBRANES</subject><subject>CELLS</subject><subject>CELLULE</subject><subject>CELULAS</subject><subject>COLD STORAGE</subject><subject>CONGELACION</subject><subject>CONGELATION</subject><subject>CONSERVACION BIOLOGICA</subject><subject>CONSERVATION BIOLOGIQUE</subject><subject>CRIOPROTECTORES</subject><subject>CRYOPROTECTANTS</subject><subject>CRYOPROTECTEUR</subject><subject>DAMAGE</subject><subject>DANOS</subject><subject>DEGAT</subject><subject>DISENO EXPERIMENTAL</subject><subject>DISPOSITIF EXPERIMENTAL</subject><subject>ESPERMATOZOO</subject><subject>EVALUACION</subject><subject>EVALUACION DEL IMPACTO</subject><subject>EVALUATION</subject><subject>EVALUATION DE L'IMPACT</subject><subject>EXPERIMENTAL DESIGN</subject><subject>FERTILIDAD</subject><subject>FERTILITE</subject><subject>FERTILITY</subject><subject>FREEZING</subject><subject>IMPACT ASSESSMENT</subject><subject>JEUNE ANIMAL</subject><subject>MEMBRANAS CELULARES</subject><subject>MEMBRANE CELLULAIRE</subject><subject>MOUVEMENT</subject><subject>MOVEMENT</subject><subject>MOVIMIENTO</subject><subject>PERMEABILIDAD</subject><subject>PERMEABILITE</subject><subject>PERMEABILITY</subject><subject>RACE (ANIMAL)</subject><subject>RAZAS (ANIMALES)</subject><subject>REPRODUCCION SEXUAL</subject><subject>REPRODUCTION SEXUEE</subject><subject>SEXUAL REPRODUCTION</subject><subject>SPERMATOZOA</subject><subject>SPERMATOZOIDE</subject><subject>STOCKAGE AU FROID</subject><subject>TEMPS</subject><subject>TIEMPO</subject><subject>TIME</subject><subject>VERRACO</subject><subject>VERRAT</subject><subject>VIABILIDAD</subject><subject>VIABILITE</subject><subject>VIABILITY</subject><subject>YOUNG ANIMALS</subject><issn>0001-7213</issn><issn>1801-7576</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><recordid>eNpdkEtLAzEUhYMoWKtrV0L-wNibxzSZpZT6gEI3unEz3GSSEpmZDEks6K93iq5cnQPnsfgIuWVwz1UtV3g0HJjUAhgAE2dkwTSwStVqfU4WACfPmbgkVzl_AHDZrMWC7Ld9GMKIJcSRRk9xilOJJVhqIiaaJ5cGLPE7Iv3MYTzQAQ-jO-VoSzhicR21ru9pjqnM-TW58Nhnd_OnS_L2uH3dPFe7_dPL5mFXWQGyVF0teeetd7rRynPDuHUelG20rRvrQYCoJXCPzgBYY0zHaqXAacWxs0KLJVn9_toUc07Ot1MKA6avlkF74tH-4zEv7n4XHmOLhxRyu3mfG_WMRkIjfgCFwF4C</recordid><startdate>20140101</startdate><enddate>20140101</enddate><creator>Mrkun, J., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creator><creator>Dolensek, T., University of Ljubljana (Slovenia). Veterinary Faculty</creator><creator>Knific, T., University of Ljubljana (Slovenia). Veterinary Faculty</creator><creator>Pislar, A., University of Ljubljana (Slovenia). Faculty of Pharmacy</creator><creator>Kosec, M., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creator><creator>Kos, J., University of Ljubljana (Slovenia). Faculty of Pharmacy</creator><creator>Zrimsek, P., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creator><scope>FBQ</scope><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>20140101</creationdate><title>Elimination of apoptotic boar spermatozoa using magnetic activated cell sorting</title><author>Mrkun, J., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses ; Dolensek, T., University of Ljubljana (Slovenia). Veterinary Faculty ; Knific, T., University of Ljubljana (Slovenia). Veterinary Faculty ; Pislar, A., University of Ljubljana (Slovenia). Faculty of Pharmacy ; Kosec, M., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses ; Kos, J., University of Ljubljana (Slovenia). Faculty of Pharmacy ; Zrimsek, P., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c304t-d542dfcfe8987f2b12cef07c98c59cf03035402faeb00cbbbd15770e872adc383</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>ALMACENAMIENTO EN FRIO</topic><topic>ANIMALES JOVENES</topic><topic>APOPTOSE</topic><topic>APOPTOSIS</topic><topic>BIOLOGICAL PRESERVATION</topic><topic>BOARS</topic><topic>BREEDS (ANIMALS)</topic><topic>CELL MEMBRANES</topic><topic>CELLS</topic><topic>CELLULE</topic><topic>CELULAS</topic><topic>COLD STORAGE</topic><topic>CONGELACION</topic><topic>CONGELATION</topic><topic>CONSERVACION BIOLOGICA</topic><topic>CONSERVATION BIOLOGIQUE</topic><topic>CRIOPROTECTORES</topic><topic>CRYOPROTECTANTS</topic><topic>CRYOPROTECTEUR</topic><topic>DAMAGE</topic><topic>DANOS</topic><topic>DEGAT</topic><topic>DISENO EXPERIMENTAL</topic><topic>DISPOSITIF EXPERIMENTAL</topic><topic>ESPERMATOZOO</topic><topic>EVALUACION</topic><topic>EVALUACION DEL IMPACTO</topic><topic>EVALUATION</topic><topic>EVALUATION DE L'IMPACT</topic><topic>EXPERIMENTAL DESIGN</topic><topic>FERTILIDAD</topic><topic>FERTILITE</topic><topic>FERTILITY</topic><topic>FREEZING</topic><topic>IMPACT ASSESSMENT</topic><topic>JEUNE ANIMAL</topic><topic>MEMBRANAS CELULARES</topic><topic>MEMBRANE CELLULAIRE</topic><topic>MOUVEMENT</topic><topic>MOVEMENT</topic><topic>MOVIMIENTO</topic><topic>PERMEABILIDAD</topic><topic>PERMEABILITE</topic><topic>PERMEABILITY</topic><topic>RACE (ANIMAL)</topic><topic>RAZAS (ANIMALES)</topic><topic>REPRODUCCION SEXUAL</topic><topic>REPRODUCTION SEXUEE</topic><topic>SEXUAL REPRODUCTION</topic><topic>SPERMATOZOA</topic><topic>SPERMATOZOIDE</topic><topic>STOCKAGE AU FROID</topic><topic>TEMPS</topic><topic>TIEMPO</topic><topic>TIME</topic><topic>VERRACO</topic><topic>VERRAT</topic><topic>VIABILIDAD</topic><topic>VIABILITE</topic><topic>VIABILITY</topic><topic>YOUNG ANIMALS</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Mrkun, J., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creatorcontrib><creatorcontrib>Dolensek, T., University of Ljubljana (Slovenia). Veterinary Faculty</creatorcontrib><creatorcontrib>Knific, T., University of Ljubljana (Slovenia). Veterinary Faculty</creatorcontrib><creatorcontrib>Pislar, A., University of Ljubljana (Slovenia). Faculty of Pharmacy</creatorcontrib><creatorcontrib>Kosec, M., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creatorcontrib><creatorcontrib>Kos, J., University of Ljubljana (Slovenia). Faculty of Pharmacy</creatorcontrib><creatorcontrib>Zrimsek, P., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</creatorcontrib><collection>AGRIS</collection><collection>CrossRef</collection><jtitle>Acta veterinaria Brno</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Mrkun, J., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</au><au>Dolensek, T., University of Ljubljana (Slovenia). Veterinary Faculty</au><au>Knific, T., University of Ljubljana (Slovenia). Veterinary Faculty</au><au>Pislar, A., University of Ljubljana (Slovenia). Faculty of Pharmacy</au><au>Kosec, M., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</au><au>Kos, J., University of Ljubljana (Slovenia). Faculty of Pharmacy</au><au>Zrimsek, P., University of Ljubljana (Slovenia). Clinic for Reproduction and Horses</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Elimination of apoptotic boar spermatozoa using magnetic activated cell sorting</atitle><jtitle>Acta veterinaria Brno</jtitle><date>2014-01-01</date><risdate>2014</risdate><volume>83</volume><issue>1</issue><spage>13</spage><epage>18</epage><pages>13-18</pages><issn>0001-7213</issn><eissn>1801-7576</eissn><abstract>One of the features of apoptosis is the externalization of phosphatidylserine which could be used to remove apoptotic cells from semen preparations. Magnetic-activated cell sorting using annexin V-conjugated microbeads which bind to phosphatidylserine could be used to enhance semen quality. Twelve boar semen samples after 3 days of liquid storage at 16ÂÂÂ17 degC were subjected to magnetic-activated cell sorting. Bound and unbound fractions and control samples were subjected to flow cytometry following the staining of spermatozoa with Annexin V conjugated with Alexa Fluor 488 and propidium iodide. Four subpopulations were obtained: live, early apoptotic live, late apoptotic, early necrotic dead and late necrotic dead. The frequency of early apoptotic and late necrotic spermatozoa was significantly higher (P less than 0.05) in bound (14.1 +/- 10.6% and 24.1 +/- 10.2%, respectively) than in unbound fractions (3.4 +/- 2.1% and 12.7 +/- 3.1%) and control (3.5 +/-+/- 1.6% and 12.0 +/- 5.0%). The lowest concentration of live spermatozoa was found in the bound fraction (10.6 +/- 8.0 %), which differed significantly (P less than 0.05) from the control. In unbound fractions there was a significantly higher concentration (P less than 0.05) of morphologically normal spermatozoa (31.8 +/- 12.6%) compared to bound ones (5.9 +/- 7.3%). A significantly (P less than 0.05) lower proportion of morphologically normal spermatozoa was observed in both fractions compared to control (67.2 +/- 17.0%). Boar spermatozoa were separated by the above method for the first time, however, the results showed this method to be inappropriate for boar semen separation under the tested conditions.</abstract><doi>10.2754/avb201483010013</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | ALMACENAMIENTO EN FRIO ANIMALES JOVENES APOPTOSE APOPTOSIS BIOLOGICAL PRESERVATION BOARS BREEDS (ANIMALS) CELL MEMBRANES CELLS CELLULE CELULAS COLD STORAGE CONGELACION CONGELATION CONSERVACION BIOLOGICA CONSERVATION BIOLOGIQUE CRIOPROTECTORES CRYOPROTECTANTS CRYOPROTECTEUR DAMAGE DANOS DEGAT DISENO EXPERIMENTAL DISPOSITIF EXPERIMENTAL ESPERMATOZOO EVALUACION EVALUACION DEL IMPACTO EVALUATION EVALUATION DE L'IMPACT EXPERIMENTAL DESIGN FERTILIDAD FERTILITE FERTILITY FREEZING IMPACT ASSESSMENT JEUNE ANIMAL MEMBRANAS CELULARES MEMBRANE CELLULAIRE MOUVEMENT MOVEMENT MOVIMIENTO PERMEABILIDAD PERMEABILITE PERMEABILITY RACE (ANIMAL) RAZAS (ANIMALES) REPRODUCCION SEXUAL REPRODUCTION SEXUEE SEXUAL REPRODUCTION SPERMATOZOA SPERMATOZOIDE STOCKAGE AU FROID TEMPS TIEMPO TIME VERRACO VERRAT VIABILIDAD VIABILITE VIABILITY YOUNG ANIMALS |
| title | Elimination of apoptotic boar spermatozoa using magnetic activated cell sorting |
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