A Shorter Equilibration Period Improves Post-Warming Outcomes after Vitrification and in Straw Dilution of In Vitro-Produced Bovine Embryos

Biology

Manel López-Béjar¹  Iris Martínez-Rodero²  Teresa Mogas²  Tania García-Martínez²  Meritxell Vendrell-Flotats²  ErikaAlina Ordóñez-León²  Carlos Olegario Hidalgo³  Marc Yeste⁴  Xabier Mendibil⁵  Joseba Esmoris⁵  Sabino Azcarate⁵ 

[1] Department of Animal Health and Anatomy, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, Spain;Department of Animal Medicine and Surgery, Autonomous University of Barcelona, ES-08193 Cerdanyola del Vallès, Spain;Department of Animal Selection and Reproduction, The Regional Agri-Food Research and Development Service of Asturias (SERIDA), ES-33394 Gijón, Spain;Department of Biology, Institute of Food and Agricultural Technology, University of Girona, ES-17004 Girona, Spain;Tekniker, Basque Research and Technology Alliance (BRTA), ES-20600 Eibar, Spain;

关键词: cow;    cryopreservation;    expanded blastocyst;    total cell number;    inner cell mass;    trophectoderm;   

DOI  :  10.3390/biology10020142

来源: DOAJ

【 摘 要 】

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 (p < 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 (p < 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 BAX, AQP3, CX43, and IFNτ gene expression between the treatments, whereas a significantly greater abundance of BCL2L1 and SOD1 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.

【 授权许可】

Unknown