Microvesicles secreted from equine amniotic- derived cells and their role in in vitro equine endometrials model

Endometris are pathologies that reduce concepon rate and increase delivery- to- concepon intervals. Many treatments have been proposed to treat or prevent endometris but in view of the embryomaternal interacon, a different approach to the treatment of endometri s could be represented by regenerave medicine. It is known that paracrine communicaon between mesenchymal stem cells and target cells exists and may involve microvesicles (MVs) as an integral component of cell- to- cell communicaon during ssue regeneraon. Based on this hypothesis, this in vitro study aims to understand the efficacy of MVs in a model of endometrial inflammaon in view of potenal applicaon in vivo. Presence and type of MVs secreted by amnioc derived cells (AMCs) was invesgated and the response of endometrial cells to MVs was studied using a dose- response curve at different concentraons (10- 20- 40- 50 × 10 6 MVs/ml) and mes (24, 48 and 72 h). Moreover, the ability of MVs to counteract in vitro inflamma on of endometrial cells induced by lipopolysaccharide (LPS) was studied through the rate of apoptosis and prolificacy, the expression of some pro- inflammatory genes such as metallopepdase (MPP) 1 and 9, IL- 1β, IL- 6 and TNF- α and the release of IL- 6, TGF- β and TNF- α. Results show that AMCs secrete MVs ranging in size from 100–200 nm. The uptake of MVs is gradual over me but peak at 72 h. MVs decrease apoptosis rate, increase prolificacy rate, down- regulate gene expression and reduce secreon of pro- inflammatory cytokines aer treatment with LPS. MiRNA 335, 146 and 26, present in these MVs, might modulate expression of the genes involved in the study.