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One-step warming does not affect the in vitro viability and cryosurvival of cryotop-vitrified donkey embryos

The objective of this study was to compare the effects of two warming protocols (three-step vs. one-step dilution) on embryo quality, post-warming embryo survival and embryo cell viability of donkey embryos vitrified by the Cryotop method. Twenty, Day 7–8, grade 1–2 donkey embryos were measured, mor...

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Published in:	Theriogenology 2020-08, Vol.152, p.47-52
Main Authors:	Bottrel, M., Hidalgo, M., Mogas, T., Pereira, B., Ortiz, I., Díaz-Jiménez, M., Consuegra, C., Morató, R., Dorado, J.
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Language:	English
Subjects:	Animals Cryopreservation - veterinary Cryotop vitrification Culture Media - chemistry Donkey embryo Embryo Culture Techniques - veterinary Embryo, Mammalian - physiology Equidae - embryology Heating One-step warming Stepwise warming Sucrose - chemistry Sucrose dilution Vitrification
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container_title	Theriogenology
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creator	Bottrel, M. Hidalgo, M. Mogas, T. Pereira, B. Ortiz, I. Díaz-Jiménez, M. Consuegra, C. Morató, R. Dorado, J.
description	The objective of this study was to compare the effects of two warming protocols (three-step vs. one-step dilution) on embryo quality, post-warming embryo survival and embryo cell viability of donkey embryos vitrified by the Cryotop method. Twenty, Day 7–8, grade 1–2 donkey embryos were measured, morphologically evaluated and vitrified using the Cryotop technique. Embryos were then randomly warmed using two different warming procedures: (i) W3 (three-step dilution; n = 11): embryos were warmed in 1 M, 0.5 M and 0 M sucrose, and (ii) W1/0.5 (one-step dilution; n = 9): embryos were warmed directly in 0.5 M sucrose. After 3 and 24 h of warming, the embryos were measured and evaluated for their morphology, developmental stage and viability (Propidium Iodide-Hoechst 33,342 dyes). Although both treatments decreased embryo quality after warming (P 0.05) were observed between protocols in terms of post-warming embryo quality, diameter and embryo survival. Greater percentages of dead cells (P
doi_str_mv	10.1016/j.theriogenology.2020.04.026
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Twenty, Day 7–8, grade 1–2 donkey embryos were measured, morphologically evaluated and vitrified using the Cryotop technique. Embryos were then randomly warmed using two different warming procedures: (i) W3 (three-step dilution; n = 11): embryos were warmed in 1 M, 0.5 M and 0 M sucrose, and (ii) W1/0.5 (one-step dilution; n = 9): embryos were warmed directly in 0.5 M sucrose. After 3 and 24 h of warming, the embryos were measured and evaluated for their morphology, developmental stage and viability (Propidium Iodide-Hoechst 33,342 dyes). Although both treatments decreased embryo quality after warming (P < 0.05), no significant differences (P > 0.05) were observed between protocols in terms of post-warming embryo quality, diameter and embryo survival. Greater percentages of dead cells (P < 0.001) were observed when embryos were warmed directly in 0.5 M sucrose (one-step dilution) when compared to the three-step protocol. The percentage of ruptured embryos was 27.3% and 0% in W3 and W1/0.5 protocols (P = 0.0893), respectively. In conclusion, warming Cryotop-vitrified donkey embryos directly in 0.5 M sucrose had no negative effects on embryo quality and post-warming embryo survival. Moreover, one-step protocol may help to prevent rupture when donkey embryos warmed directly in 0.5 M sucrose. These results observed in vitro must be verified by embryo transfer. •One-step sucrose dilution was tested for warming of Cryotop-vitrified donkey embryos.•Warming directly in 0.5 M sucrose did not affect embryo quality and survival.•One-step protocol may help to prevent rupture of embryos.</description><identifier>ISSN: 0093-691X</identifier><identifier>EISSN: 1879-3231</identifier><identifier>DOI: 10.1016/j.theriogenology.2020.04.026</identifier><identifier>PMID: 32361488</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Animals ; Cryopreservation - veterinary ; Cryotop vitrification ; Culture Media - chemistry ; Donkey embryo ; Embryo Culture Techniques - veterinary ; Embryo, Mammalian - physiology ; Equidae - embryology ; Heating ; One-step warming ; Stepwise warming ; Sucrose - chemistry ; Sucrose dilution ; Vitrification</subject><ispartof>Theriogenology, 2020-08, Vol.152, p.47-52</ispartof><rights>2020 Elsevier Inc.</rights><rights>Copyright © 2020 Elsevier Inc. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c440t-67009d8d1cfb9c2786d360e900c04e3e8f80cb6e14670ae547b7447fbdfd39593</citedby><cites>FETCH-LOGICAL-c440t-67009d8d1cfb9c2786d360e900c04e3e8f80cb6e14670ae547b7447fbdfd39593</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32361488$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bottrel, M.</creatorcontrib><creatorcontrib>Hidalgo, M.</creatorcontrib><creatorcontrib>Mogas, T.</creatorcontrib><creatorcontrib>Pereira, B.</creatorcontrib><creatorcontrib>Ortiz, I.</creatorcontrib><creatorcontrib>Díaz-Jiménez, M.</creatorcontrib><creatorcontrib>Consuegra, C.</creatorcontrib><creatorcontrib>Morató, R.</creatorcontrib><creatorcontrib>Dorado, J.</creatorcontrib><title>One-step warming does not affect the in vitro viability and cryosurvival of cryotop-vitrified donkey embryos</title><title>Theriogenology</title><addtitle>Theriogenology</addtitle><description>The objective of this study was to compare the effects of two warming protocols (three-step vs. one-step dilution) on embryo quality, post-warming embryo survival and embryo cell viability of donkey embryos vitrified by the Cryotop method. Twenty, Day 7–8, grade 1–2 donkey embryos were measured, morphologically evaluated and vitrified using the Cryotop technique. Embryos were then randomly warmed using two different warming procedures: (i) W3 (three-step dilution; n = 11): embryos were warmed in 1 M, 0.5 M and 0 M sucrose, and (ii) W1/0.5 (one-step dilution; n = 9): embryos were warmed directly in 0.5 M sucrose. After 3 and 24 h of warming, the embryos were measured and evaluated for their morphology, developmental stage and viability (Propidium Iodide-Hoechst 33,342 dyes). Although both treatments decreased embryo quality after warming (P < 0.05), no significant differences (P > 0.05) were observed between protocols in terms of post-warming embryo quality, diameter and embryo survival. Greater percentages of dead cells (P < 0.001) were observed when embryos were warmed directly in 0.5 M sucrose (one-step dilution) when compared to the three-step protocol. The percentage of ruptured embryos was 27.3% and 0% in W3 and W1/0.5 protocols (P = 0.0893), respectively. In conclusion, warming Cryotop-vitrified donkey embryos directly in 0.5 M sucrose had no negative effects on embryo quality and post-warming embryo survival. Moreover, one-step protocol may help to prevent rupture when donkey embryos warmed directly in 0.5 M sucrose. These results observed in vitro must be verified by embryo transfer. •One-step sucrose dilution was tested for warming of Cryotop-vitrified donkey embryos.•Warming directly in 0.5 M sucrose did not affect embryo quality and survival.•One-step protocol may help to prevent rupture of embryos.</description><subject>Animals</subject><subject>Cryopreservation - veterinary</subject><subject>Cryotop vitrification</subject><subject>Culture Media - chemistry</subject><subject>Donkey embryo</subject><subject>Embryo Culture Techniques - veterinary</subject><subject>Embryo, Mammalian - physiology</subject><subject>Equidae - embryology</subject><subject>Heating</subject><subject>One-step warming</subject><subject>Stepwise warming</subject><subject>Sucrose - chemistry</subject><subject>Sucrose dilution</subject><subject>Vitrification</subject><issn>0093-691X</issn><issn>1879-3231</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><recordid>eNqNkMFu1DAQhi0EokvhFZAPHLgkjGOvk0hcUEWhUqVeQOJmOfZ48ZLYi50NytvwLDwZXrZF4sZpZM03848_Ql4xqBkw-WZfz18x-bjDEMe4W-sGGqhB1NDIR2TDuraveMPZY7IB6Hkle_blgjzLeQ8AXEr2lFyUvmSi6zZkugtY5RkP9IdOkw87aiNmGuJMtXNoZlrSqA-_fi5-TpEuXg9-9PNKdbDUpDXmY1r8okca3Z_3HA_VifXOoy3bwjdcKU7DCX1Onjg9ZnxxXy_J5-v3n64-Vrd3H26u3t1WRgiYK9mWy21nmXFDb5q2k5ZLwB7AgECOnevADBKZKKTGrWiHVojWDdZZ3m97fklen_ceUvx-xDyryWeD46gDxmNWDe87JsUW2oK-PaMmxZwTOnVIftJpVQzUSbjaq3-Fq5NwBUIV4WX85X3ScZjQ_h1-MFyA6zOA5b-Lx6Sy8RgMWp-KXmWj_7-k38ajngw</recordid><startdate>202008</startdate><enddate>202008</enddate><creator>Bottrel, M.</creator><creator>Hidalgo, M.</creator><creator>Mogas, T.</creator><creator>Pereira, B.</creator><creator>Ortiz, I.</creator><creator>Díaz-Jiménez, M.</creator><creator>Consuegra, C.</creator><creator>Morató, R.</creator><creator>Dorado, J.</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>202008</creationdate><title>One-step warming does not affect the in vitro viability and cryosurvival of cryotop-vitrified donkey embryos</title><author>Bottrel, M. ; Hidalgo, M. ; Mogas, T. ; Pereira, B. ; Ortiz, I. ; Díaz-Jiménez, M. ; Consuegra, C. ; Morató, R. ; Dorado, J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c440t-67009d8d1cfb9c2786d360e900c04e3e8f80cb6e14670ae547b7447fbdfd39593</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Animals</topic><topic>Cryopreservation - veterinary</topic><topic>Cryotop vitrification</topic><topic>Culture Media - chemistry</topic><topic>Donkey embryo</topic><topic>Embryo Culture Techniques - veterinary</topic><topic>Embryo, Mammalian - physiology</topic><topic>Equidae - embryology</topic><topic>Heating</topic><topic>One-step warming</topic><topic>Stepwise warming</topic><topic>Sucrose - chemistry</topic><topic>Sucrose dilution</topic><topic>Vitrification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Bottrel, M.</creatorcontrib><creatorcontrib>Hidalgo, M.</creatorcontrib><creatorcontrib>Mogas, T.</creatorcontrib><creatorcontrib>Pereira, B.</creatorcontrib><creatorcontrib>Ortiz, I.</creatorcontrib><creatorcontrib>Díaz-Jiménez, M.</creatorcontrib><creatorcontrib>Consuegra, C.</creatorcontrib><creatorcontrib>Morató, R.</creatorcontrib><creatorcontrib>Dorado, J.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Theriogenology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bottrel, M.</au><au>Hidalgo, M.</au><au>Mogas, T.</au><au>Pereira, B.</au><au>Ortiz, I.</au><au>Díaz-Jiménez, M.</au><au>Consuegra, C.</au><au>Morató, R.</au><au>Dorado, J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>One-step warming does not affect the in vitro viability and cryosurvival of cryotop-vitrified donkey embryos</atitle><jtitle>Theriogenology</jtitle><addtitle>Theriogenology</addtitle><date>2020-08</date><risdate>2020</risdate><volume>152</volume><spage>47</spage><epage>52</epage><pages>47-52</pages><issn>0093-691X</issn><eissn>1879-3231</eissn><abstract>The objective of this study was to compare the effects of two warming protocols (three-step vs. one-step dilution) on embryo quality, post-warming embryo survival and embryo cell viability of donkey embryos vitrified by the Cryotop method. 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subjects	Animals Cryopreservation - veterinary Cryotop vitrification Culture Media - chemistry Donkey embryo Embryo Culture Techniques - veterinary Embryo, Mammalian - physiology Equidae - embryology Heating One-step warming Stepwise warming Sucrose - chemistry Sucrose dilution Vitrification
title	One-step warming does not affect the in vitro viability and cryosurvival of cryotop-vitrified donkey embryos
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