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当前位置: 首页 > 文章 > Extracellular vesicles from oviductal and uterine fluids supplementation in sequential in vitro culture improves bovine embryo quality 畜牧与生物技术杂志(英文版) 2022,13 (1)
Position: Home > Articles > Extracellular vesicles from oviductal and uterine fluids supplementation in sequential in vitro culture improves bovine embryo quality Journal of Animal Science and Biotechnology 2022,13 (1)

Extracellular vesicles from oviductal and uterine fluids supplementation in sequential in vitro culture improves bovine embryo quality

作  者：
Cláudia Lima Verde Leal;K. Cañón-Beltrán;Y. N. Cajas;Meriem Hamdi;Aracelli Yaryes;María Gemma Millán de la Blanca;P. Beltrán-Breña;Rosane Mazzarella;Juliano Coelho da Silveira;Alfonso Gutiérrez‐Adán;Esther Sánchez-Pardo González;Dimitrios Rizo
单  位：
Facultad de Ciencias Agrarias y Ambientales, Programa de Medicina Veterinaria, Fundación Universitaria Juan de Castellanos, Tunja, Colombia;Department of Animal Reproduction, National Center Institute for Agriculture and Food Research and Technology (CSIC-INIA), Madrid, Spai; Department of Animal Reproduction, National Center Institute for Agriculture and Food Research and Technology (CSIC-INIA), Madrid, Spain;Department of Animal Reproduction, National Center Institute for Agriculture and Food Research and Technology (CSIC-INIA), Madrid, Spain;Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo (FZEA-USP), Pirassununga, Brazil; Laboratorio de Biotecnología de la Reproducción Animal, Facultad de Ciencias Agropecuarias, Universidad de Cuenca (UC), Cuenca, Ecuador;Department of Anatomy and Embryology, Veterinary Faculty-Universidad Complutense de Madrid (UCM), Madrid, Spain
关键词：
Cattle;Cryopreservation;Embryo development;Exosomes;Lipid metabolism;Oviduct;Uterus;miRNA
摘  要：
In vitro production of bovine embryos is a well-established technology, but the in vitro culture (IVC) system still warrants improvements, especially regarding embryo quality. This study aimed to evaluate the effect of extracellular vesicles (EVs) isolated from oviductal (OF) and uterine fluid (UF) in sequential IVC on the development and quality of bovine embryos. Zygotes were cultured in SOF supplemented with either BSA or EVs-depleted fetal calf serum (dFCS) in the presence (BSA-EV and dFCS-EV) or absence of EVs from OF (D1 to D4) and UF (D5 to D8), mimicking in vivo conditions. EVs from oviducts (early luteal phase) and uterine horns (mid-luteal phase) from slaughtered heifers were isolated by size exclusion chromatography. Blastocyst rate was recorded on days 7-8 and their quality was assessed based on lipid contents, mitochondrial activity and total cell numbers, as well as survival rate after vitrification. Relative mRNA abundance for lipid metabolism-related transcripts and levels of phosphorylated hormone-sensitive lipase (pHSL) proteins were also determined. Additionally, the expression levels of 383 miRNA in OF- and UF-EVs were assessed by qRT-PCR.Blastocyst yield was lower (P < 0.05) in BSA treatments compared with dFCS treatments. Survival rates after vitrification/warming were improved in dFCS-EVs (P < 0.05). EVs increased (P < 0.05) blastocysts total cell number in dFCS-EV and BSA-EV compared with respective controls (dFCS and BSA), while lipid content was decreased in dFCS-EV (P < 0.05) and mitochondrial activity did not change (P > 0.05). Lipid metabolism transcripts were affected by EVs and showed interaction with type of protein source in medium (PPARGC1B, LDLR, CD36, FASN and PNPLA2, P < 0.05). Levels of pHSL were lower in dFCS (P < 0.05). Twenty miRNA were differentially expressed between OF- and UF-EVs and only bta-miR-148b was increased in OF-EVs (P < 0.05).Mimicking physiological conditions using EVs from OF and UF in sequential IVC does not affect embryo development but improves blastocyst quality regarding survival rate after vitrification/warming, total cell number, lipid content, and relative changes in expression of lipid metabolism transcripts and lipase activation. Finally, EVs miRNA contents may contribute to the observed effects.

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