Oviduct epithelial cell‑derived extracellular vesicles promote the developmental competence of IVF porcine embryos

Assisted reproductive technology has increased the efficiency of animal reproduction. However, polyspermy is a significant limitation of porcine in vitro fertilization (IVF). Therefore, reducing the polyspermy rate and improving monospermic embryos is crucial. Recent studies have reported that oviductal fluid, along with its contents of extracellular vesicles (EVs), enhanced the fertilization process and supported embryo development. Consequently, the present study investigated the effects of porcine oviduct epithelial cells (OEC-EVs) on sperm-oocyte interactions during porcine IVF and evaluated in vitro embryo developmental competence outcomes. During IVF embryo development, the cleavage rate was significantly higher in the group treated with 50 ng/ml OEC-EVs compared with the control group (67.6±2.5 vs. 57.3±1.9; P<0.05). Furthermore, the OEC-EV group had significantly more embryos (16.4±1.2 vs. 10.2±0.8; P<0.05), and the polyspermy rate significantly decreased (32.9±2.5 vs. 43.8±3.1; P<0.05) compared with that of the control group. Additionally, the fluorescence intensities of cortical granules (3.56±0.47 vs. 2.15±0.24; P<0.05) and active mitochondria (8.14±0.34 vs. 5.96±0.38; P<0.05) were significantly higher in the OEC-EV group compared with those in the control group. In conclusion, OEC-EV adsorption and penetration crosstalk between sperm and oocytes was observed. OEC-EV treatment was demonstrated to significantly improve the concentration and distribution of cortical granules in oocytes. Furthermore, OEC-EVs also increased oocyte mitochondrial activity, reduced polyspermy and increased the IVF success rate.

[1]  Jongki Cho,et al.  Oviduct Epithelial Cell-Derived Extracellular Vesicles Improve Porcine Trophoblast Outgrowth , 2022, Veterinary sciences.

[2]  Jongki Cho,et al.  Oviduct epithelial cells‐derived extracellular vesicles improve preimplantation developmental competence of in vitro produced porcine parthenogenetic and cloned embryos , 2021, Molecular reproduction and development.

[3]  M. Ericsson,et al.  Extracellular Vesicles Protect the Neonatal Lung from Hyperoxic Injury Through the Epigenetic and Transcriptomic Reprogramming of Myeloid Cells. , 2021, American journal of respiratory and critical care medicine.

[4]  P. Mermillod,et al.  Editorial: Biofluid Extracellular Vesicles and Their Involvement in Animal Reproductive Physiology , 2021, Frontiers in Veterinary Science.

[5]  D. Kang,et al.  Modified Spirulina maxima Pectin Nanoparticles Improve the Developmental Competence of In Vitro Matured Porcine Oocytes , 2021, Animals : an open access journal from MDPI.

[6]  Soo-Hyun Park,et al.  Luteolin Orchestrates Porcine Oocyte Meiotic Progression by Maintaining Organelle Dynamics Under Oxidative Stress , 2021, Frontiers in Cell and Developmental Biology.

[7]  U. Chandran,et al.  Microvesicles Transfer Mitochondria and Increase Mitochondrial Function in Brain Endothelial Cells , 2021, bioRxiv.

[8]  F. Perecin,et al.  Changes in Oviductal Cells and Small Extracellular Vesicles miRNAs in Pregnant Cows , 2021, Frontiers in Veterinary Science.

[9]  H. Kaneko,et al.  Excess polyspermy reduces the ability of porcine oocytes to promote male pronuclear formation after in vitro fertilization , 2021, Animal science journal = Nihon chikusan Gakkaiho.

[10]  Hyo-Jin Park,et al.  Anti-oxidative effects of exogenous ganglioside GD1a and GT1b on embryonic developmental competence in pigs , 2020, Journal of Animal Reproduction and Biotechnology.

[11]  C. Matás,et al.  Involvement of nitric oxide during in vitro oocyte maturation, sperm capacitation and in vitro fertilization in pig. , 2020, Research in veterinary science.

[12]  W. Winuthayanon,et al.  Extracellular Vesicles and the Oviduct Function , 2020, International journal of molecular sciences.

[13]  M. Kulus,et al.  Cortical Granule Distribution and Expression Pattern of Genes Regulating Cellular Component Size, Morphogenesis, and Potential to Differentiation are Related to Oocyte Developmental Competence and Maturational Capacity In Vivo and In Vitro , 2020, Genes.

[14]  P. Mermillod,et al.  Porcine oviductal extracellular vesicles interact with gametes and regulate sperm motility and survival. , 2020, Theriogenology.

[15]  S. Bauersachs,et al.  Extracellular vesicles: Multi-signal messengers in the gametes/embryo-oviduct cross-talk. , 2020, Theriogenology.

[16]  F. Perecin,et al.  Extracellular Vesicles Mediated Early Embryo–Maternal Interactions , 2020, International journal of molecular sciences.

[17]  H. Kaneko,et al.  Selection based on morphological features of porcine embryos produced by in vitro fertilization: Timing of early cleavages and the effect of polyspermy , 2020, Animal science journal = Nihon chikusan Gakkaiho.

[18]  F. Silvestris,et al.  Extracellular Vesicles and Epigenetic Modifications Are Hallmarks of Melanoma Progression , 2019, International journal of molecular sciences.

[19]  P. Mermillod,et al.  Oviduct fluid extracellular vesicles regulate polyspermy during porcine invitro fertilisation. , 2019, Reproduction, fertility, and development.

[20]  Wenjun Zhou,et al.  Melatonin enhances mitochondrial biogenesis and protects against rotenone‐induced mitochondrial deficiency in early porcine embryos , 2019, Journal of pineal research.

[21]  Z. Ding,et al.  The toxic effects of Fluorene‐9‐bisphenol on porcine oocyte in vitro maturation , 2019, Environmental toxicology.

[22]  J. Gadea,et al.  Pig in vitro fertilization: Where are we and where do we go? , 2019, Theriogenology.

[23]  M. Noonan,et al.  Oviductal extracellular vesicles interact with the spermatozoon’s head and mid-piece and improves its motility and fertilizing ability in the domestic cat , 2019, Scientific Reports.

[24]  J. Roca,et al.  High pre-freezing sperm dilution improves monospermy without affecting the penetration rate in porcine IVF. , 2019, Theriogenology.

[25]  S. Bauersachs,et al.  Extracellular Vesicles in the Oviduct: Progress, Challenges and Implications for the Reproductive Success , 2019, Bioengineering.

[26]  Dylan T Burnette,et al.  Reassessment of Exosome Composition , 2019, Cell.

[27]  J. Simonsen.  Pitfalls associated with lipophilic fluorophore staining of extracellular vesicles for uptake studies , 2019, Journal of extracellular vesicles.

[28]  W. Geldenhuys,et al.  Extracellular Vesicles Secreted in Response to Cytokine Exposure Increase Mitochondrial Oxygen Consumption in Recipient Cells , 2019, Front. Cell. Neurosci..

[29]  A. Kumaresan,et al.  Effect of bovine oviductal fluid on motility, tyrosine phosphorylation, and acrosome reaction in cryopreserved bull spermatozoa. , 2019, Theriogenology.

[30]  Clotilde Théry,et al.  Specificities of secretion and uptake of exosomes and other extracellular vesicles for cell-to-cell communication , 2019, Nature Cell Biology.

[31]  Z. Fereshteh,et al.  Oviductal extracellular vesicles (oviductosomes, OVS) are conserved in humans: murine OVS play a pivotal role in sperm capacitation and fertility , 2018, Molecular human reproduction.

[32]  D. Yellon,et al.  Confounding factors in vesicle uptake studies using fluorescent lipophilic membrane dyes , 2017, Journal of extracellular vesicles.

[33]  P. Mermillod,et al.  Oviduct extracellular vesicles protein content and their role during oviduct-embryo cross-talk. , 2017, Reproduction.

[34]  Dagmar Wachten,et al.  Intracellular activation of ovastacin mediates pre-fertilization hardening of the zona pellucida , 2017, Molecular human reproduction.

[35]  M. Gorospe,et al.  RNA in extracellular vesicles , 2017, Wiley interdisciplinary reviews. RNA.

[36]  Janet Iwasa,et al.  Mechanisms of action and regulation of ATP-dependent chromatin-remodelling complexes , 2017, Nature Reviews Molecular Cell Biology.

[37]  W. Pang,et al.  Boar seminal plasma exosomes maintain sperm function by infiltrating into the sperm membrane , 2016, Oncotarget.

[38]  J. Souza-Fabjan,et al.  Combination of oviduct fluid and heparin to improve monospermic zygotes production during porcine in vitro fertilization. , 2016, Theriogenology.

[39]  A. Baccarelli,et al.  Extracellular vesicles: roles in gamete maturation, fertilization and embryo implantation. , 2015, Human reproduction update.

[40]  E. Seli,et al.  Oocyte mitochondrial function and reproduction , 2015, Current opinion in obstetrics & gynecology.

[41]  Artur Lichtenberg,et al.  An Innovative Method for Exosome Quantification and Size Measurement , 2015, Journal of visualized experiments : JoVE.

[42]  Byeong-chun Lee,et al.  Improvement of cloned embryos development by co-culturing with parthenotes: a possible role of exosomes/microvesicles for embryos paracrine communication. , 2014, Cellular reprogramming.

[43]  E. Strehler,et al.  Expression and Secretion of Plasma Membrane Ca2+-ATPase 4a (PMCA4a) during Murine Estrus: Association with Oviductal Exosomes and Uptake in Sperm , 2013, PloS one.

[44]  R. Lloyd,et al.  Effects of porcine pre-ovulatory oviductal fluid on boar sperm function. , 2010, Theriogenology.

[45]  Marcos J. Araúzo-Bravo,et al.  Chromatin-Remodeling Components of the BAF Complex Facilitate Reprogramming , 2010, Cell.

[46]  J. Roca,et al.  Advances in swine in vitro embryo production technologies. , 2010, Reproduction in domestic animals = Zuchthygiene.

[47]  M. Avilés,et al.  What controls polyspermy in mammals, the oviduct or the oocyte? , 2009, Biological reviews of the Cambridge Philosophical Society.

[48]  Yi Wu,et al.  Abnormal changes in mitochondria, lipid droplets, ATP and glutathione content, and Ca(2+) release after electro-activation contribute to poor developmental competence of porcine oocyte during in vitro ageing. , 2009, Reproduction, fertility, and development.

[49]  M. Avilés,et al.  Oviduct-specific glycoprotein and heparin modulate sperm–zona pellucida interaction during fertilization and contribute to the control of polyspermy , 2008, Proceedings of the National Academy of Sciences.

[50]  S. Antonini,et al.  Porcine embryonic stem cells: Facts, challenges and hopes. , 2007, Theriogenology.

[51]  I. Saadeldin,et al.  Role of Exosomes in Biological Communication Systems , 2021 .

[52]  I. Saadeldin.  Extracellular Vesicles Mediate the Embryonic-Maternal Paracrine Communication , 2020 .

[53]  Jongki Cho,et al.  The Interplay Between Exosomes and Spermatozoa , 2020 .

[54]  M. Hirata,et al.  Journal of Reproduction and Development , 2019 .

[55]  W. Winuthayanon,et al.  Oviduct: roles in fertilization and early embryo development. , 2017, The Journal of endocrinology.

[56]  M. Avilés,et al.  The oviduct: A key organ for the success of early reproductive events , 2015 .

[57]  C. Grupen.  The evolution of porcine embryo in vitro production. , 2014, Theriogenology.

[58]  D. Beebe,et al.  Developments in in vitro technologies for swine embryo production. , 2004, Reproduction, fertility, and development.

[59]  Stem Cells and Cloning: Advances and Applications Dovepress , 2022 .