Enrichment of thawed boar spermatozoa with an intact membrane using Magnetic Activated Cell Sorting
Not all boar sperm samples survive cryopreservation well. A method of eliminating damaged sperm might enable more cryopreserved boar semen to be used for pig breeding. In this study we investigated the use of Magnetic Activated Cell sorting (MACS) to eliminate damaged sperm from thawed boar semen sa...
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| Veröffentlicht in: | Animal reproduction science 2024-06, Vol.265, p.107493-107493, Article 107493 |
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| creator | Johannisson, Anders Morrell, Jane M. Wallgren, Margareta |
| description | Not all boar sperm samples survive cryopreservation well. A method of eliminating damaged sperm might enable more cryopreserved boar semen to be used for pig breeding. In this study we investigated the use of Magnetic Activated Cell sorting (MACS) to eliminate damaged sperm from thawed boar semen samples. The thawed samples were mixed with Dead cell removal particles and were applied to the column in a SuperMACS II. Different fractions were collected: Original sample (O), Flow-through (FT), and Eluate (E). Sperm membrane integrity, mitochondrial membrane potential and reactive oxygen species were evaluated by flow cytometry after staining with SYBR 14 and propidium iodide, or 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethylbenzimidazolylcarbocyanine iodide, or hydroethidine and dichlorodihydrofluorescein diacetate, respectively. The FT samples had increased membrane integrity, a greater proportion of sperm with high mitochondrial membrane potential and a greater proportion of sperm negative for hydrogen peroxide than O samples (P |
| doi_str_mv | 10.1016/j.anireprosci.2024.107493 |
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[Display omitted]
•Boar sperm quality deteriorated during freezing and thawing.•After MACS, one third of the original sperm number appeared in flow-through fraction.•Sperm quality was enhanced in the flow-through fraction compared to the original.•Eluate had worse sperm quality than flow-through fraction or original sample.</description><identifier>ISSN: 0378-4320</identifier><identifier>ISSN: 1873-2232</identifier><identifier>EISSN: 1873-2232</identifier><identifier>DOI: 10.1016/j.anireprosci.2024.107493</identifier><identifier>PMID: 38701639</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>animal reproduction ; Apoptosis ; boars ; Clinical Science ; cryopreservation ; Cryopreserved boar semen ; flow cytometry ; hydrogen peroxide ; iodides ; Klinisk vetenskap ; MACS ; magnetism ; membrane potential ; mitochondrial membrane ; propidium ; Reactive Oxygen Species ; Reproductive biotechnologies ; semen ; Sperm membrane damage ; sperm quality</subject><ispartof>Animal reproduction science, 2024-06, Vol.265, p.107493-107493, Article 107493</ispartof><rights>2024 The Authors</rights><rights>Copyright © 2024. Published by Elsevier B.V.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c430t-d6e166c60aa66665f4396495546af060baabd990ea4197fc3c6b960a1db7a55f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0378432024000848$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>230,314,550,776,780,881,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38701639$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-538279$$DView record from Swedish Publication Index$$Hfree_for_read</backlink><backlink>$$Uhttps://res.slu.se/id/publ/131696$$DView record from Swedish Publication Index$$Hfree_for_read</backlink></links><search><creatorcontrib>Johannisson, Anders</creatorcontrib><creatorcontrib>Morrell, Jane M.</creatorcontrib><creatorcontrib>Wallgren, Margareta</creatorcontrib><creatorcontrib>Sveriges lantbruksuniversitet</creatorcontrib><title>Enrichment of thawed boar spermatozoa with an intact membrane using Magnetic Activated Cell Sorting</title><title>Animal reproduction science</title><addtitle>Anim Reprod Sci</addtitle><description>Not all boar sperm samples survive cryopreservation well. A method of eliminating damaged sperm might enable more cryopreserved boar semen to be used for pig breeding. In this study we investigated the use of Magnetic Activated Cell sorting (MACS) to eliminate damaged sperm from thawed boar semen samples. The thawed samples were mixed with Dead cell removal particles and were applied to the column in a SuperMACS II. Different fractions were collected: Original sample (O), Flow-through (FT), and Eluate (E). Sperm membrane integrity, mitochondrial membrane potential and reactive oxygen species were evaluated by flow cytometry after staining with SYBR 14 and propidium iodide, or 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethylbenzimidazolylcarbocyanine iodide, or hydroethidine and dichlorodihydrofluorescein diacetate, respectively. The FT samples had increased membrane integrity, a greater proportion of sperm with high mitochondrial membrane potential and a greater proportion of sperm negative for hydrogen peroxide than O samples (P<0.0001), which in turn had increased membrane integrity than E samples (P <0.0001). However, differences were seen between boars. The FT samples had increased values of live, superoxide positive sperm than O samples (P <0.0001) and O samples had greater values than E samples (P <0.0001), while there was no effect of boar. Sperm quality was best in the FT fraction, comprising approximately 32% of the sperm sample. In conclusion, although there were differences between boars, MACS separation can improve sperm quality in thawed semen samples. It would be interesting to see if this improvement is reflected in fertility outcomes.
[Display omitted]
•Boar sperm quality deteriorated during freezing and thawing.•After MACS, one third of the original sperm number appeared in flow-through fraction.•Sperm quality was enhanced in the flow-through fraction compared to the original.•Eluate had worse sperm quality than flow-through fraction or original sample.</description><subject>animal reproduction</subject><subject>Apoptosis</subject><subject>boars</subject><subject>Clinical Science</subject><subject>cryopreservation</subject><subject>Cryopreserved boar semen</subject><subject>flow cytometry</subject><subject>hydrogen peroxide</subject><subject>iodides</subject><subject>Klinisk vetenskap</subject><subject>MACS</subject><subject>magnetism</subject><subject>membrane potential</subject><subject>mitochondrial membrane</subject><subject>propidium</subject><subject>Reactive Oxygen Species</subject><subject>Reproductive biotechnologies</subject><subject>semen</subject><subject>Sperm membrane damage</subject><subject>sperm quality</subject><issn>0378-4320</issn><issn>1873-2232</issn><issn>1873-2232</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>D8T</sourceid><recordid>eNqNkV-PEyEUxYnRuHX1Kxh888GpMDBMeWzqupqs8cE_r-TC3GlpOkMFZhv99Esz68YXo7yQwO-cC-cQ8oqzJWdcvd0vYfQRjzEk55c1q2U5b6UWj8iCr1pR1bWoH5MFE-2qkqJmF-RZSnvGWKuUfkouxKotPkIviLsao3e7AcdMQ0_zDk7YURsg0nTEOEAOvwLQk887CiP1YwaX6YCDjTAinZIft_QTbEfM3tG1y_4WcnHY4OFAv4SYy_1z8qSHQ8IX9_sl-fb-6uvmQ3Xz-frjZn1TOSlYrjqFXCmnGIAqq-ml0ErqppEKeqaYBbCd1gxBct32TjhldaF5Z1toml5ckuXsm054nKw5Rj9A_GkCeJMOk4V43kxCwwVXWhXBm78K3vnvaxPi1kyTacSqbnXBX894Cf7HhCmbwSdXPlqSCFMyghewaWVT_xtlDdOSSX1G9Yy6UmeK2D88gzNzrtvszR91m3PdZq67aF_ej5nsgN2D8ne_BdjMAJbcbz2WBJzH0WFXDF02XfD_MeYOX-zC_g</recordid><startdate>20240601</startdate><enddate>20240601</enddate><creator>Johannisson, Anders</creator><creator>Morrell, Jane M.</creator><creator>Wallgren, Margareta</creator><general>Elsevier B.V</general><scope>6I.</scope><scope>AAFTH</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7S9</scope><scope>L.6</scope><scope>ACNBI</scope><scope>ADTPV</scope><scope>AOWAS</scope><scope>D8T</scope><scope>DF2</scope><scope>ZZAVC</scope></search><sort><creationdate>20240601</creationdate><title>Enrichment of thawed boar spermatozoa with an intact membrane using Magnetic Activated Cell Sorting</title><author>Johannisson, Anders ; Morrell, Jane M. ; Wallgren, Margareta</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c430t-d6e166c60aa66665f4396495546af060baabd990ea4197fc3c6b960a1db7a55f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>animal reproduction</topic><topic>Apoptosis</topic><topic>boars</topic><topic>Clinical Science</topic><topic>cryopreservation</topic><topic>Cryopreserved boar semen</topic><topic>flow cytometry</topic><topic>hydrogen peroxide</topic><topic>iodides</topic><topic>Klinisk vetenskap</topic><topic>MACS</topic><topic>magnetism</topic><topic>membrane potential</topic><topic>mitochondrial membrane</topic><topic>propidium</topic><topic>Reactive Oxygen Species</topic><topic>Reproductive biotechnologies</topic><topic>semen</topic><topic>Sperm membrane damage</topic><topic>sperm quality</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Johannisson, Anders</creatorcontrib><creatorcontrib>Morrell, Jane M.</creatorcontrib><creatorcontrib>Wallgren, Margareta</creatorcontrib><creatorcontrib>Sveriges lantbruksuniversitet</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>SWEPUB Uppsala universitet full text</collection><collection>SwePub</collection><collection>SwePub Articles</collection><collection>SWEPUB Freely available online</collection><collection>SWEPUB Uppsala universitet</collection><collection>SwePub Articles full text</collection><jtitle>Animal reproduction science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Johannisson, Anders</au><au>Morrell, Jane M.</au><au>Wallgren, Margareta</au><aucorp>Sveriges lantbruksuniversitet</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enrichment of thawed boar spermatozoa with an intact membrane using Magnetic Activated Cell Sorting</atitle><jtitle>Animal reproduction science</jtitle><addtitle>Anim Reprod Sci</addtitle><date>2024-06-01</date><risdate>2024</risdate><volume>265</volume><spage>107493</spage><epage>107493</epage><pages>107493-107493</pages><artnum>107493</artnum><issn>0378-4320</issn><issn>1873-2232</issn><eissn>1873-2232</eissn><abstract>Not all boar sperm samples survive cryopreservation well. A method of eliminating damaged sperm might enable more cryopreserved boar semen to be used for pig breeding. In this study we investigated the use of Magnetic Activated Cell sorting (MACS) to eliminate damaged sperm from thawed boar semen samples. The thawed samples were mixed with Dead cell removal particles and were applied to the column in a SuperMACS II. Different fractions were collected: Original sample (O), Flow-through (FT), and Eluate (E). Sperm membrane integrity, mitochondrial membrane potential and reactive oxygen species were evaluated by flow cytometry after staining with SYBR 14 and propidium iodide, or 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethylbenzimidazolylcarbocyanine iodide, or hydroethidine and dichlorodihydrofluorescein diacetate, respectively. The FT samples had increased membrane integrity, a greater proportion of sperm with high mitochondrial membrane potential and a greater proportion of sperm negative for hydrogen peroxide than O samples (P<0.0001), which in turn had increased membrane integrity than E samples (P <0.0001). However, differences were seen between boars. The FT samples had increased values of live, superoxide positive sperm than O samples (P <0.0001) and O samples had greater values than E samples (P <0.0001), while there was no effect of boar. Sperm quality was best in the FT fraction, comprising approximately 32% of the sperm sample. In conclusion, although there were differences between boars, MACS separation can improve sperm quality in thawed semen samples. It would be interesting to see if this improvement is reflected in fertility outcomes.
[Display omitted]
•Boar sperm quality deteriorated during freezing and thawing.•After MACS, one third of the original sperm number appeared in flow-through fraction.•Sperm quality was enhanced in the flow-through fraction compared to the original.•Eluate had worse sperm quality than flow-through fraction or original sample.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>38701639</pmid><doi>10.1016/j.anireprosci.2024.107493</doi><tpages>1</tpages><oa>free_for_read</oa></addata></record> |
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| source | Elsevier ScienceDirect Journals; SWEPUB Freely available online |
| subjects | animal reproduction Apoptosis boars Clinical Science cryopreservation Cryopreserved boar semen flow cytometry hydrogen peroxide iodides Klinisk vetenskap MACS magnetism membrane potential mitochondrial membrane propidium Reactive Oxygen Species Reproductive biotechnologies semen Sperm membrane damage sperm quality |
| title | Enrichment of thawed boar spermatozoa with an intact membrane using Magnetic Activated Cell Sorting |
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