A simple and rapid flow cytometry-based assay to identify a competent embryo prior to embryo transfer

Pállinger, Éva; Bognár, Zita; Bódis, József; Csabai, Timea; Farkas, Nelli; Gödöny, Krisztina; Várnagy, Ákos; Buzás, Edit I.; Szekeres-Barthó, Júlia

doi:10.1038/srep39927

Cited by 39 publications

(36 citation statements)

References 37 publications

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“…EVs are produced by all types of cells, and because they transport different kinds of molecules from one cell to the other, they can be considered as means of intercellular communication, and as such, might be considered as candidates for conveying the signal from the embryo to the mother. Earlier, we showed that in vitro cultured human embryos produce detectable numbers of EVs 4 , therefore, it seemed plausible, that these structures might be involved in the communication between the embryo and the endometrium during implantation.…”

Section: Introductionmentioning

confidence: 96%

PIBF+ extracellular vesicles from mouse embryos affect IL-10 production by CD8+ cells

Pállinger

Bognár²,

Bogdan

et al. 2018

Sci Rep

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Earlier evidence suggests, that the embryo signals to the maternal immune system. Extracellular vesicles (EVs) are produced by all types of cells, and because they transport different kinds of molecules from one cell to the other, they can be considered as means of intercellular communication. The aim of this work was to test, whether the embryo is able to produce sufficient amounts of EVs to alter the function of peripheral lymphocytes. Embryo-derived EVs were identified by their Annexin V biding capacity, and sensitivity to Triton X dependent lysis, using flow cytometry. Transmission electron microscopy was used to detect EVs at the implantation site. Progesterone-induced blocking factor (PIBF) expression in embryo-derived EVs was demonstrated with immuno-electron microscopy. The % of IL-10 + murine lymphocytes was determined by flow cytometry. EVs were present in embryo culture media, but not in empty media. Mouse embryo-derived EVs adhere to the surface of both CD4+ and CD8+ murine peripheral T lymphocytes, partly, via phosphatidylserine binding. The number of IL-10+ murine peripheral CD8+ cells increases in the presence of embryo-derived EVS, and this effect is counteracted by pre-treatment of EVs with an anti-PIBF antibody, suggesting that the embryo communicates with the maternal immune system via EVs.

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Section: Introductionmentioning

confidence: 96%

PIBF+ extracellular vesicles from mouse embryos affect IL-10 production by CD8+ cells

Pállinger

Bognár²,

Bogdan

et al. 2018

Sci Rep

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“…A recent study reported the presence of DNA in embryo-derived EVs by flow cytometry, associating higher concentrations of DNA-containing EVs with higher rates of implantation failure after embryo transfer. Nevertheless, this may be explained by the fact that these DNA-containing EVs were mainly apoptotic bodies, thus reflecting poor embryo quality [55]. To our knowledge, there are still no works addressing the potential diagnostic value of embryo derived EVs for the detection of DNA mutations/polymorphisms in the cells of origin, but some works in different biofluids, such as blood, urine, or seminal fluid, have done so, highlighting EVs as potential biomarkers for pathological mutations [56].…”

Section: Discussionmentioning

confidence: 99%

Identification and Characterization of Extracellular Vesicles and Its DNA Cargo Secreted During Murine Embryo Development

Simon

Bolumar

Amadoz

et al. 2020

Genes

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Extracellular vesicles (EVs) are known to transport DNA, but their implications in embryonic implantation are unknown. The aim of this study was to investigate EVs production and secretion by preimplantation embryos and assess their DNA cargo. Murine oocytes and embryos were obtained from six- to eight-week-old females, cultured until E4.5 and analyzed using transmission electron microscopy to examine EVs production. EVs were isolated from E4.5-day conditioned media and quantified by nanoparticle tracking analysis, characterized by immunogold, and their DNA cargo sequenced. Multivesicular bodies were observed in murine oocytes and preimplantation embryos together with the secretion of EVs to the blastocoel cavity and blastocyst spent medium. Embryo-derived EVs showed variable electron-densities and sizes (20–500 nm) and total concentrations of 1.74 × 107 ± 2.60 × 106 particles/mL. Embryo secreted EVs were positive for CD63 and ARF6. DNA cargo sequencing demonstrated no differences in DNA between apoptotic bodies or smaller EVs, although they showed significant gene enrichment compared to control medium. The analysis of sequences uniquely mapping the murine genome revealed that DNA contained in EVs showed higher representation of embryo genome than vesicle-free DNA. Murine blastocysts secrete EVs containing genome-wide sequences of DNA to the medium, reinforcing the relevance of studying these vesicles and their cargo in the preimplantation moment, where secreted DNA may help the assessment of the embryo previous to implantation.

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“…We have previously demonstrated EV in culture media of in vitro cultured human embryos [71]; thus it seemed plausible that these structures might be involved in communication between the embryo and the endometrium during implantation.…”

Section: The Embryo Communicates With the Maternal Immune System Via mentioning

confidence: 99%

The Role of Progesterone in Feto-Maternal Immunological Cross Talk

Szekeres-Barthó

2018

Med Princ Pract

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This review aims to provide a brief historical overview of the feto-maternal immunological relationship, which profoundly influences the outcome of pregnancy. The initial question posed in the 1950s by Medawar [Symp Soc Exp Biol. 1953; 7: 320–338] was based on the assumption that the maternal immune system recognizes the fetus as an allograft. Indeed, based on the association between HLA-matching and spontaneous miscarriage, it became obvious that immunological recognition of pregnancy is required for a successful gestation. The restricted expression of polymorphic HLA antigens on the trophoblast, together with the presence of nonpolymorphic MHC products, excludes recognition by both T and NK cells of trophoblast-presented antigens; however, γδ T cells, which constitute the majority of decidual T cells, are likely candidates. Indeed, a high number of activated, progesterone receptor-expressing γδ T cells are present in the peripheral blood of healthy pregnant women and, in the presence of progesterone, these cells secrete an immunomodulatory protein called progesterone-induced blocking factor (PIBF). As early as in the peri-implantation period, the embryo communicates with the maternal immune system via PIBF containing extracellular vesicles. PIBF contributes to the dominance of Th2-type reactivity which characterizes normal pregnancy by inducing increased production of Th2 cytokines. The high expression of this molecule in the decidua might be one of the reasons for the low cytotoxic activity of decidual NK cells.

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A simple and rapid flow cytometry-based assay to identify a competent embryo prior to embryo transfer

Cited by 39 publications

References 37 publications

PIBF+ extracellular vesicles from mouse embryos affect IL-10 production by CD8+ cells

PIBF+ extracellular vesicles from mouse embryos affect IL-10 production by CD8+ cells

Identification and Characterization of Extracellular Vesicles and Its DNA Cargo Secreted During Murine Embryo Development

The Role of Progesterone in Feto-Maternal Immunological Cross Talk

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