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Extracellular vesicles (EVs) have become important in embryo-maternal communication during early development. The aim of this study was to evaluate the effect of an in vitro system on early bidirectional embryo-maternal communication mediated by EVs. For this purpose, two experiments were performed: one to evaluate the effect of embryonic EVs on maternal cells and the second to determine the effect of maternal EVs on early embryonic development. For the first in vitro (IVP) and in vivo (IVV) experiments, bovine blastocysts were selected and individually cultured for 48 h to collect embryonic EVs secreted during days 7-9 of embryonic development. Embryonic EVs were added to the medium of in vitro-cultured bovine endometrial cells to evaluate their effect on the expression pattern of genes associated with endometrial function and response to interferon tau (IFNT). Non-classical interferon-stimulated genes (ISGs) were only induced by in vitro-derived embryos. In the second experiment, EVs released by endometrial cells cultured in vitro (EVC) and collected from uterine fluid (EV-UF) of cows in the early luteal phase were added to the culture medium of bovine embryos produced in vitro during days 5-9 of development. The effect of maternal in vitro or in vivo-derived EVs differs in the quality of bovine embryos produced in vitro during the pre-implantation period. The expression of IFNT in bovine embryos is increased by the effect of EV-UF treatment. Additionally, EV-UF treatment induces a sustained increase in diameter during embryonic development and a tendency towards a greater number of expanded and hatched blastocysts. However, some genes related to embryo quality are induced by EVC treatment., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Mesenchymal stem cells (MSC) are modern tools in regenerative therapies of humans and animals owed to their immunomodulatory properties, which are activated in a pro-inflammatory environment. Different preconditioning strategies had been devised to enhance the immunomodulatory properties of MSC. In this research, we evaluated the immunological attributes of equine adipose MSC (eAMSC) before and after preconditioning in vitro with prostaglandin E 2 (PGE 2 ), substance P (SP), their combination and IFNγ. PGE 2 /SP was the best combination to keep or enhance the mesodermal lineage differentiation of eAMSC. Alongside with this, preconditioning of eMSC with PGE 2 and SP did not affect expression of stemness MSC surface phenotype: CD90 + , CD44 + , MHC class I + , MHC class II - and CD45 - , assessed by cytometry. Both naïve and preconditioned eAMSC expressed genes related with immune properties, such as MHC-I, PTGES, IL6, IL1A, TNFα and IL8 assessed by qPCR. Only TNFα was under expressed in treated cells, while the other markers were either overexpressed or not changed. In no cases MHC-II expression was detected. The antiproliferative effect of preconditioned eAMSC exposed to activated peripheral blood mononuclear cells (PBMC) showed that SP treatment significantly inhibited proliferation of LPS stimulated PBMC. When eAMSC were stimulated with Poly I:C, all the treatments significantly inhibited proliferation of stimulated PBMC (p < 0.05). Direct contact (coculture) between the preconditioned eAMSC and PBMC, induced a shift of significantly more (CD4/CD25/FOXP3) + T-regulatory PBMC than naïve eAMSC. In the experiments of this research, we investigated the secreted proteomic profile of naïve and preconditioned eAMSC, 42 up-regulated and 40 down-regulated proteins were found in the proteomic assay. Our proteomic data revealed profound changes in the secretory pattern of MSC exposed to different treatments, compared to naïve eAMSC as well as among treatments. In overall, compared to naïve cells, the protein profile of preconditioned cells resembled the mesenchymal-epithelial transition (MET). Here we showed that the combined use of PGE 2 and SP provoked in overall the highest expression of anti-inflammatory markers as well as lead to an increased acquisition of a T-regulatory phenotype in preconditioned eAMSC without affecting their "stemness"., (Copyright © 2020 Elsevier B.V. All rights reserved.)