Oviduct epithelial cells‐derived extracellular vesicles improve preimplantation developmental competence of in vitro produced porcine parthenogenetic and cloned embryos

Abstract

Extracellular vesicles (EVs) carry bioactive cargoes involved in the early preimplantation development. This study investigated the effects of EVs obtained from an oviductal epithelial cell (OEC) conditioned medium on the developmental competence of in parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) porcine embryos. The OEC‐EV‐treated group showed significant increases in blastocyst formation and hatching rates compared to the control group (40.8% ± 2.2% and 20.1% ± 2.1% vs. 24.9% ± 2.0% and 5.3% ± 1.1%; p < 0.05), respectively. The 7 day OEC‐EVs treatment group significantly increased blastocyst formation rate than the 3 day and 0 day‐groups (45.0 ± 0.8 vs. 33.0 ± 0.7 and 26.7 ± 0.5; p < 0.05), respectively. SCNT revealed that the OEC‐EV increased blastocyst formation rate compared to that of oviductal fluid EVs (OF‐EVs) (35.4% ± 1.4% vs. 29.3% ± 1.3%; p < 0.05). Reactive oxygen species levels, apoptosis, and blastocyst lipid content were significantly decreased in the OEC‐EVs group compared with the control group. OEC‐EV group showed a significantly decreased BAX and increased BCL2, SOD1, POU5F1, SOX2, NANOG, GATA6, PNPLA2, LIPE, and MGLL gene expression than the control group (p < 0.05). In conclusion, OEC‐EVs supplementation in embryo culture media improved the quality of porcine embryos, potentially helping porcine‐cloned embryonic development possibly through transfer of messenger RNA and proteins to the early embryos.

Oviductal Extracellular Vesicles Enhance Porcine In Vitro Embryo Development by Modulating the Embryonic Transcriptome

“…Similarly, Fang et al [ 56 ] reported that oEV obtained from in vitro oviductal epithelial cell-conditioned media improved the development competence of parthenogenetic and somatic nuclear transfer porcine embryos. In contrast to our results, supplementation for 7 days provided better blastocyst rates than compared to three days with in vitro obtained oEVs.…”

“…The enhanced embryo development, survival, and the downregulation of SIRT1 in EVs-treated embryos suggests the possibility of a post-transcriptional downregulation of SIRT1 in EV-treated embryos by miRNAs [ 83 ]. In this way, SIRT1 in embryos could be potentially targeted by miR-21-5p or miR-9-5p present in bovine oEVs and in porcine oEVs [ 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 ]. In the present study, although SIRT1 was not ...…”

“…This leads to the hypothesis that the downregulation of HERPUD2 in EV-treated embryos, potentially due to miRNA regulation (e.g., mir-7-5p in poEVs), might protect the embryos from stressful IVC conditions and enhance their development in vitro. In this line, Fang et al [ 56 ] suggested that oEVs from in vitro origin could minimize the oxidative damage associated to in vitro environment.…”

Messenger roles of extracellular vesicles during fertilization of gametes, development and implantation: Recent advances

“…The oviduct is the site of zygote birth as well as preembryonic development. Current research indicates that O-EVs can improve embryo blastocyst formation rate, embryo survival rate, embryo hatching rate, and post-embryo transfer birth rate ( Almiñana et al, 2017 ; Qu et al, 2019 ; Fang et al, 2022b ; de Alcântara-Neto et al, 2022 ). Furthermore, O-EVs can improve embryo development after cryopreservation and survival rate after vitrification ( Lopera-Vasquez et al, 2017 ; Leal et al, 2022 ; Sidrat et al, 2022 ).…”

Isolation, characterization, proteome, miRNAome, and the embryotrophic effects of chicken egg yolk nanovesicles (vitellovesicles)

“…In vitro generation of porcine parthenogenetic embryos. Electric parthenogenetic activation of in vitro matured porcine oocytes was used to generate porcine embryos as showed in previous reports 16,17,28 . Briefly, ovaries were collected from a local slaughterhouse in Daejeon city, and cumulus-oocyte complexes (COCs) were aspirated.…”