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Characterization of extracellular vesicles produced by single human embryos at early stages of development

Tankov, S., Lavrits, A., Lättekivi, F., Krjutskov, K., Sikut, A., Kõks, S., Andronowska, A., Salumets, A. and Fazeli, A. (2018) Characterization of extracellular vesicles produced by single human embryos at early stages of development. Journal of Extracellular Vesicles, 7 (Supp. 1). p. 37.

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Background: Extracellular vesicles (EVs) are recognized as potent vehicles for intercellular communication. To date, there is little information available regarding the role of EVs during the early stages of human embryonic development. The aim of this study was to develop techniques for the recovery of EVs secreted by a single human embryo in an in vitro culture system. The EVs were characterized according to size, concentration and electrical surface properties (zeta potential), in order to understand the role of EVs production in human embryos for determination of their quality at early stages of development. Methods: Human embryos were produced by in vitro fertilization (IVF) for 24 h in fertilization medium, cultured individually for 48 h (3 days) in cleavage medium and additionally 48 h in blastocyst medium (day 5). Conditioned media, at days 3 and 5 post-IVF, was collected and EVs were isolated using a series of centrifugations and size-exclusion chromatography. The size, concentration and zeta potential of EVs were characterized using a nanoparticle tracking analysis. Results: Using this method of isolation, we were able to collect and characterize EVs produced by a single human embryo. Analysis confirmed the presence of EVs at early stages of development, with the concentration of EVs being higher in early blastocysts (day 5), as compared to 4-8 cell-stage embryos (day 3). Moreover, already at day 3, we were able to discriminate between embryos that were properly developing and those that were later visually determined as degrading at day 5. The data indicates that embryos following normal development at day 3 but degrading at later stages (day 5) were producing significantly higher number of EVs (with size range of 100-160 nm) compared with those developing properly at day 3 and later progressing to early blastocysts at day 5. Summary/conclusion: In conclusion, we have developed a sensitive protocol for the isolation of EVs from human embryos cultured individually. We have demonstrated that human embryos secrete EVs in varying amounts and sizes during the early stages of their development. Further investigations are needed to establish EV characteristics of early human embryo as a quality marker for human clinical embryology.

Item Type: Journal Article
Publisher: Taylor and Francis Open
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