A Shorter Equilibration Period Improves Post-Warming Outcomes after Vitrification and in Straw Dilution of In Vitro-Produced Bovine Embryos
This study was designed to the optimize vitrification and in-straw warming protocol of in vitro-produced bovine embryos by comparing two different equilibration periods, short equilibrium (SE: 3 min) and long equilibrium (LE: 12 min). Outcomes recorded in vitrified day seven (D7) and day eight (D8)...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-02-01
|
| Series: | Biology |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2079-7737/10/2/142 |
| _version_ | 1797411583030198272 |
|---|---|
| author | Iris Martínez-Rodero Tania García-Martínez Erika Alina Ordóñez-León Meritxell Vendrell-Flotats Carlos Olegario Hidalgo Joseba Esmoris Xabier Mendibil Sabino Azcarate Manel López-Béjar Marc Yeste Teresa Mogas |
| author_facet | Iris Martínez-Rodero Tania García-Martínez Erika Alina Ordóñez-León Meritxell Vendrell-Flotats Carlos Olegario Hidalgo Joseba Esmoris Xabier Mendibil Sabino Azcarate Manel López-Béjar Marc Yeste Teresa Mogas |
| author_sort | Iris Martínez-Rodero |
| collection | DOAJ |
| description | This study was designed to the optimize vitrification and in-straw warming protocol of in vitro-produced bovine embryos by comparing two different equilibration periods, short equilibrium (SE: 3 min) and long equilibrium (LE: 12 min). Outcomes recorded in vitrified day seven (D7) and day eight (D8) expanded blastocysts were survival and hatching rates, cell counts, apoptosis rate, and gene expression. While survival rates at 3 and 24 h post-warming were reduced (<i>p</i> < 0.05) after vitrification, the hatching rates of D7 embryos vitrified after SE were similar to the rates recorded in fresh non-vitrified blastocysts. The hatching rates of vitrified D8 blastocysts were lower (<i>p</i> < 0.05) than of fresh controls regardless of treatment. Total cell count, and inner cell mass and trophectoderm cell counts were similar in hatched D7 blastocysts vitrified after SE and fresh blastocysts, while vitrified D8 blastocysts yielded lower values regardless of treatment. The apoptosis rate was significantly higher in both treatment groups compared to fresh controls, although rates were lower for SE than LE. No differences emerged in <i>BAX</i>, <i>AQP3</i>, <i>CX43,</i> and <i>IFNτ</i> gene expression between the treatments, whereas a significantly greater abundance of <i>BCL2L1</i> and <i>SOD1</i> transcripts was observed in blastocysts vitrified after SE. A shorter equilibration vitrification protocol was found to improve post-warming outcomes and time efficiency after in-straw warming/dilution. |
| first_indexed | 2024-03-09T04:47:13Z |
| format | Article |
| id | doaj.art-b83d7873618d4731964c82f6faa1be28 |
| institution | Directory Open Access Journal |
| issn | 2079-7737 |
| language | English |
| last_indexed | 2024-03-09T04:47:13Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biology |
| spelling | doaj.art-b83d7873618d4731964c82f6faa1be282023-12-03T13:14:12ZengMDPI AGBiology2079-77372021-02-0110214210.3390/biology10020142A Shorter Equilibration Period Improves Post-Warming Outcomes after Vitrification and in Straw Dilution of In Vitro-Produced Bovine EmbryosIris Martínez-Rodero0Tania García-Martínez1Erika Alina Ordóñez-León2Meritxell Vendrell-Flotats3Carlos Olegario Hidalgo4Joseba Esmoris5Xabier Mendibil6Sabino Azcarate7Manel López-Béjar8Marc Yeste9Teresa Mogas10Department of Animal Medicine and Surgery, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, SpainDepartment of Animal Medicine and Surgery, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, SpainDepartment of Animal Medicine and Surgery, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, SpainDepartment of Animal Medicine and Surgery, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, SpainDepartment of Animal Selection and Reproduction, The Regional Agri-Food Research and Development Service of Asturias (SERIDA), ES-33394 Gijón, SpainTekniker, Basque Research and Technology Alliance (BRTA), ES-20600 Eibar, SpainTekniker, Basque Research and Technology Alliance (BRTA), ES-20600 Eibar, SpainTekniker, Basque Research and Technology Alliance (BRTA), ES-20600 Eibar, SpainDepartment of Animal Health and Anatomy, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, SpainDepartment of Biology, Institute of Food and Agricultural Technology, University of Girona, ES-17004 Girona, SpainDepartment of Animal Medicine and Surgery, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, SpainThis study was designed to the optimize vitrification and in-straw warming protocol of in vitro-produced bovine embryos by comparing two different equilibration periods, short equilibrium (SE: 3 min) and long equilibrium (LE: 12 min). Outcomes recorded in vitrified day seven (D7) and day eight (D8) expanded blastocysts were survival and hatching rates, cell counts, apoptosis rate, and gene expression. While survival rates at 3 and 24 h post-warming were reduced (<i>p</i> < 0.05) after vitrification, the hatching rates of D7 embryos vitrified after SE were similar to the rates recorded in fresh non-vitrified blastocysts. The hatching rates of vitrified D8 blastocysts were lower (<i>p</i> < 0.05) than of fresh controls regardless of treatment. Total cell count, and inner cell mass and trophectoderm cell counts were similar in hatched D7 blastocysts vitrified after SE and fresh blastocysts, while vitrified D8 blastocysts yielded lower values regardless of treatment. The apoptosis rate was significantly higher in both treatment groups compared to fresh controls, although rates were lower for SE than LE. No differences emerged in <i>BAX</i>, <i>AQP3</i>, <i>CX43,</i> and <i>IFNτ</i> gene expression between the treatments, whereas a significantly greater abundance of <i>BCL2L1</i> and <i>SOD1</i> transcripts was observed in blastocysts vitrified after SE. A shorter equilibration vitrification protocol was found to improve post-warming outcomes and time efficiency after in-straw warming/dilution.https://www.mdpi.com/2079-7737/10/2/142cowcryopreservationexpanded blastocysttotal cell numberinner cell masstrophectoderm |
| spellingShingle | Iris Martínez-Rodero Tania García-Martínez Erika Alina Ordóñez-León Meritxell Vendrell-Flotats Carlos Olegario Hidalgo Joseba Esmoris Xabier Mendibil Sabino Azcarate Manel López-Béjar Marc Yeste Teresa Mogas A Shorter Equilibration Period Improves Post-Warming Outcomes after Vitrification and in Straw Dilution of In Vitro-Produced Bovine Embryos Biology cow cryopreservation expanded blastocyst total cell number inner cell mass trophectoderm |
| title | A Shorter Equilibration Period Improves Post-Warming Outcomes after Vitrification and in Straw Dilution of In Vitro-Produced Bovine Embryos |
| title_full | A Shorter Equilibration Period Improves Post-Warming Outcomes after Vitrification and in Straw Dilution of In Vitro-Produced Bovine Embryos |
| title_fullStr | A Shorter Equilibration Period Improves Post-Warming Outcomes after Vitrification and in Straw Dilution of In Vitro-Produced Bovine Embryos |
| title_full_unstemmed | A Shorter Equilibration Period Improves Post-Warming Outcomes after Vitrification and in Straw Dilution of In Vitro-Produced Bovine Embryos |
| title_short | A Shorter Equilibration Period Improves Post-Warming Outcomes after Vitrification and in Straw Dilution of In Vitro-Produced Bovine Embryos |
| title_sort | shorter equilibration period improves post warming outcomes after vitrification and in straw dilution of in vitro produced bovine embryos |
| topic | cow cryopreservation expanded blastocyst total cell number inner cell mass trophectoderm |
| url | https://www.mdpi.com/2079-7737/10/2/142 |
| work_keys_str_mv | AT irismartinezrodero ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT taniagarciamartinez ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT erikaalinaordonezleon ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT meritxellvendrellflotats ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT carlosolegariohidalgo ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT josebaesmoris ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT xabiermendibil ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT sabinoazcarate ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT manellopezbejar ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT marcyeste ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT teresamogas ashorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT irismartinezrodero shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT taniagarciamartinez shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT erikaalinaordonezleon shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT meritxellvendrellflotats shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT carlosolegariohidalgo shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT josebaesmoris shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT xabiermendibil shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT sabinoazcarate shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT manellopezbejar shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT marcyeste shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos AT teresamogas shorterequilibrationperiodimprovespostwarmingoutcomesaftervitrificationandinstrawdilutionofinvitroproducedbovineembryos |