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

作　　者：Cláudia Lima Verde Leal Karina Canón-Beltrán Yulia N.Cajas Meriem Hamdi Aracelli Yaryes María Gemma Millán de la Blanca Paula Beltrán-Brena Rosane Mazzarella Juliano Coelho da Silveira Alfonso Gutiérrez-Adán Encina M González Dimitrios Rizos 

机构地区：[1]Department of Animal Reproduction,National Center Institute for Agriculture and Food Research and Technology(CSIC-INIA),28040 Madrid,Spain. [2]Departamento de Medicina Veterinaria,Faculdade de Zootecnia e Engenharia de Alimentos,Universidade de Sao Paulo(FZEA-USP),Pirassununga,Brazil. [3]Facultad de Ciencias Agrarias y Ambientales,Programa de Medicina Veterinaria,Fundacion Universitaria Juan de Castellanos,Tunja,Colombia. [4]Laboratorio de Biotecnologia de la Reproduccion Animal,Facultad de Ciencias Agropecuarias,Universidad de Cuenca(UC),EC010205 Cuenca,Ecuador. [5]Department of Anatomy and Embryology,Veterinary Faculty-Universidad Complutense de Madrid(UCM),Madrid,Spain.

出　　处：《Journal of Animal Science and Biotechnology》2023年第1期114-133,共20页畜牧与生物技术杂志（英文版）

基　　金：supported by the Spanish Ministry of Science and Innovation (PID2019-111641RB-I00 to DR, and RTI2018-093548-B-I00 to AGA);Sáo Paulo Research Foundation,Brazil (FAPESP;#2017/20339-3 and CNPqBrazil 304276/2018-9 to CLVL;#2019/04981-2 to RM;2014/22887-0 and 2015/21829-9 to JCDS);Minciencias-Colombia Postdoctoral Fellowship (848-2019) to KC-B;Secretaria de Educación Superior,Ciencia y Tecnología e Innovación (SENESCYT-Ecuador) to YNC.

摘　　要：Background: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 hormonesensitive lipase(pHSL)proteins were also determined.Additionally,the expression levels of 383 miRNA in OF-and UF-EVs were assessed by qRT-PCR.Results: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 dFCSEV(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).Conclusions: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 vitr

关 键 词：CATTLE CRYOPRESERVATION Embryo development EXOSOMES Lipid metabolism miRNAs OVIDUCT UTERUS 

分 类 号：S823[农业科学—畜牧学]