To evaluate the effect of conditioned media (CM) and Extracellular Vesicles (EVs) derived from bovine oviduct epithelial cell (BOEC) lines on developmental capacity zygotes quality embryos produced in vitro, presumptive were cultured under specific conditions. In experiment 1, either monolayers BOEC extended culture (E), together with fresh suspension cells, or BOEC-CM E-monolayers. 2, EVs isolated characterized (150–200 nm) by Nanosight® electron microscopy. Zygotes presence 3x105EVs/mL,...

The aim of this study was to evaluate the effect extracellular vesicles (EV) from oviductal fluid (OF), either ampulla or isthmus, on development and quality in vitro -cultured bovine embryos. Zygotes were cultured synthetic oviduct (SOF + 3 mg/mL BSA) without calf serum (C− group), presence × 10 5 EV/mL ampullary isthmic OF at 1 4 g (10 K) (100 K), compared with SOF 5% FCS (C+ group). OF-EV size concentration assessed by electron microscopy nanotracking analysis system. Embryo recorded Days...

Abstract Background Extracellular vesicles (EVs) present in oviductal (OF) and uterine fluid (UF) have been shown to enhance bovine embryo quality during vitro culture by reducing lipid contents modulating metabolism-related genes (LMGs), while also influencing cell proliferation, suggesting their involvement on the regulation of different biological pathways. The signaling pathways related differentiation, metabolism is crucial for early development can determine success or failure...

To evaluate the effect of bovine oviductal fluid (OF) supplementation during in vitro culture embryos on their development and quality, vitro-produced zygotes were cultured synthetic (SOF; negative control; C-) supplemented with OF or 5% fetal calf serum (positive C+). Embryo was recorded Days 7-9 after insemination blastocyst quality assessed through cryotolerance, differential cell counting inner mass trophectoderm, gene expression. added to medium at concentrations ranging from 0.625%...

Abstract Background In vitro production of bovine embryos is a well-established technology, but the in culture (IVC) system still warrants improvements, especially regarding embryo quality. This study aimed to evaluate effect extracellular vesicles (EVs) isolated from oviductal (OF) and uterine fluid (UF) sequential IVC on development quality embryos. Zygotes were cultured SOF supplemented with either BSA or EVs-depleted fetal calf serum (dFCS) presence (BSA-EV dFCS-EV) absence EVs OF (D1...

In order to mimic the maternal oviductal environment, we evaluated effect of fluid (OF) and/or uterine (UF) supplementation on in vitro embryo development and quality. vitro-produced zygotes were cultured with 1.25% OF from Day 1 4 after insemination (OF group), followed by UF 9 (OF+UF group) or only (UF group). Control groups presence synthetic oviduct (SOF) supplemented 3mgmL-1 bovine serum albumin (BSA) 5% fetal calf (FCS). Supplementation culture medium (both at 1.25%) supported (Day...

Signaling components of bone morphogenetic proteins (BMPs) are expressed in an anatomically and temporally regulated fashion bovine oviduct. However, a local response this signaling to the presence embryo has yet be elucidated. The aim present study was evaluate if early embryo-oviduct interaction induces changes gene expression BMP components. For purpose, we used vitro co-culture system investigate between oviductal epithelial cells (BOEC) from isthmus region with embryos during two...

During the transit through oviduct, early embryo initiates an extensive DNA methylation reprogramming of its genome. Given that these epigenetic modifications are susceptible to environmental factors, components present in oviductal milieu could affect marks developing embryo. The aim this study was examine if culture bovine embryos with fluid (OF) can induce changes at specific genomic regions resulting blastocysts. In vitro produced zygotes were cultured medium 3 mg/mL serum albumin (BSA)...

Contents Antioxidants have been widely used during in vitro production to decrease the negative effect of reactive oxygen species. It was reported that complex resveratrol–methyl β‐cyclodextrin ( RV ‐ CD ) improves resveratrol's stability and bioavailability increases its antioxidant activity. This study evaluates oocyte maturation IVM or embryo culture IVC on developmental competence quantitative changes gene expression important genes. In experiment 1, added media level, development...

Maternal aging-associated reduction of oocyte viability is a common feature in mammals, but more research needed to counteract this process.In women, the first aging phenotype appears with decline reproductive function, and follicle number gradually decreases from menarche menopause.Cows can be used as model early human embryonic development because both species share very high degree similarity during selection, cleavage, blastocyst formation.Recently, it has been proposed that main driver...

Extracellular vesicles (EVs) found in various biological fluids and particularly reproductive fluids, have gained considerable attention for their possible role cell- to- cell communication. Among, the different bioactive molecules cargos of EVs, MicroRNAs (miRNAs) are emerging as promising diagnostic biomarkers with high clinical potential. Aiming to understand roles EVs bovine tract, we intended characterize profile oviduct uterine (OF-EVs, UF-EVs) miRNA across estrous cycle. Nanoparticle...

During its journey through the oviduct, bovine embryo may induce transcriptomic and metabolic responses, via direct or indirect contact, from oviduct epithelial cells (BOECs). An in vitro model using polyester mesh was established, allowing study of local contact during 48 h between a BOEC monolayer early embryos (2- 8-cell stage) their respective conditioned media (CM). The response to assessed by analyzing transcript abundance SMAD6, TDGF1, ROCK1, ROCK2, SOCS3, PRELP AGR3 selected previous...

The increasing use of in vitro embryo production (IVP) followed by transfer (ET), alongside with cryopreservation embryos, has risen concerns regarding the possible altered pregnancy rates, calving or even neonatal mortality. One hypotheses for these alterations is current culture conditions IVP. In an attempt to better mimic physiological milieu, embryos were produced female reproductive fluids (RF) as supplements medium, and another group supplemented bovine serum albumin (BSA) control....

This study aimed to examine the local embryo effect on transcriptomic response of epithelial cells oviduct in vivo. Fifteen heifers were synchronized and artificially inseminated a standing heat. All slaughtered Day 2.5 after oestrus. The oviducts from 13 animals isolated, trimmed free tissue divided between ampulla/isthmus. ipsilateral isthmus was into smaller sections (2 cm). Each section sequentially flushed until located (4/13) then opened scraped longitudinally obtain cells. Cells...

Abstract In contrast to other domestic mammals, the embryo-derived signal(s) leading maternal recognition of pregnancy (MRP) are still unknow in mare. We hypothesize that these embryonic signals could be packed into uterine extracellular vesicles (uEVs), acting as multi-signal messengers between conceptus and tract, contributing MRP. To unveil signals, RNA protein cargos uEVs isolated from lavages collected pregnant mares (P; day 10, 11, 12 13 after ovulation) cyclic control (C; 10 were...

Abstract Background Equine embryos exhibit an unusual pattern of glucose tolerance in vitro and are currently cultured hyperglycaemic conditions. Objective Our main objective was to analyse the effect different concentrations on vitro‐produced equine embryo development quality. Study design Experiments comparing vivo produced embryos. Methods Oocytes (n = 641) were collected from post‐mortem ovaries, matured fertilised by intracytoplasmic sperm injection (ICSI). Embryo culture divided Day 0...

Assisted reproductive technologies impact transcriptome and epigenome of embryos can result in long-term phenotypic consequences. Whole-genome DNA methylation profiles from individual bovine blastocysts vivo- vitro-derived (using three sources protein: fluids, blood serum albumin) were generated. The vitro culture on was analyzed, sex-specific differences at blastocyst stage uncovered. In vivo showed the highest levels (29.5%), close to those produced with serum, whilst fluids or albumin...

The aim of this study was to determine the effect maternal–embryonic asynchrony in reproductive tract (oviduct and uterus) on subsequent embryo development cattle. Fifty Day 1invitro-produced zygotes were transferred endoscopically into oviduct ipsilateral corpus luteum heifers (n=40) that either synchronous with embryos (Day 1 after ovulation) or asynchronous ahead 3 ovulation). A subset killed a commercial abattoir 3, 6 14 days transfer. Location within tract, developmental stage quality...

Contents Ascorbic acid (AC) used as antioxidant in embryo culture is very sensitive and degrades unavoidably aqueous solution. Methyl‐β‐cyclodextrin (CD) improved the stability of AC solution to elevated temperature, light, humidity oxidation. The aim this study was evaluate effect complex AC‐CD during vitro maturation (IVM) or (IVC) on oocyte developmental competence subsequent development quality. (100 µM) added IVM media, level were examined. Matured oocytes, their cumulus cells produced...

Although fertilization and early embryonic development take place in the oviduct, consequences of tubal fluid supplementation during vitro embryo culture have not been explored. The aim this study was to evaluate effect bovine oviducal (bOF) embryos on their quality. bOF aspirated from oviducts slaughtered heifers luteal phase. In vitro-produced zygotes were cultured SOF (C–; n = 927) or + 5% FCS (C+; 872) (n ~900/group) at different concentrations (0.62, 1.25, 2.5, 5, 10, 25%) 10...