Clinical Trial: Isolation and Characterization of the Extracellular Vesicles Secreted by the Human Endometrium
Study Status: Recruiting
Recruit Status: Recruiting
Study Type: Observational
Official Title: Isolation and Characterization of the Extracellular Vesicles Secreted by the Human Endometrium to the Endometrial Fluid
Brief Summary: The main aim of this study is to describe the morphology, size distributions and specific markers of the different vesicles populations present in the human endometrial fluid. To further characterize the vesicles populations, contents of these membrane compartments (i.e.: DNA, RNA, proteins, lipids and other small metabolites) will be assessed by using mass spectrometry and next generation sequencing techniques. The objective here is to relate a specific vesicles composition to the moment of the cycle, to a normal physiological endometrial function and to the transmission of functionalities from the mother to the embryo.
Detailed Summary:
In the last decades, the emission of membrane enclosed compartments, more commonly regarded as extracellular vesicles, has been established as a new mechanism of communication either between cells of a complex organism, between unicellular organisms or between unicellular organism and host. These vesicles have been described in all body fluids and are currently classified into three types according to the mechanism by which they are formed: exosomes, with sizes ranging from 30 to 150 nm (formed intracellularly); microvesicles, between 100 and 1000 nm (formed from the cell membrane) and apoptotic bodies, with sizes greater than 1 µm (produced by cells undergoing programmed cell death).
Endometrial fluid is a viscous liquid present in the uterine cavity originating from the different cell types that form the uterus and serum exudates. It offers a precise view of the context in which the embryo implants and can be retrieved by a minimally invasive technique.
The main objective of this pilot study is to isolate and characterize the extracellular vesicles secreted by the endometrium to the endometrial fluid, being this the first time that this fact is being described. It is intended to standardize a method for the isolation of the different vesicles populations present within the endometrial fluid (i.e.: apoptotic bodies, microvesicles and exosomes), based on serial differential centrifugations and filtration. Morphological characterization of the isolated vesicles will be done at three levels: study of the external morphology by transmission electron microscopy, characterization of the different vesicles populations surface markers by Western Blot and analysis of the vesicles populations size distributions by Nanoparticle Tracking Analysis.
The second part of the study is
Sponsor: Igenomix
Current Primary Outcome:
- Morphological characterization of the extracellular vesicles [ Time Frame: 3 months ]
Assessment of the morphology of the extracellular vesicles using transmision electron microscopy by either ultrathin cuts and positive staining or deposition and positive staining techniques.
The aim is to differentiate between vesicle populations thanks to their external aspects and size.
- Size distribution profiles characterization of the extracellular vesicles [ Time Frame: 45 days ]
Analysis of the size distribution of the vesicles contained in the different extracellular vesicles populations by using two Nanoparticle Tracking Analysis techniques: ZetaSizer Nano and NanoSight300.
The expected outcome is a set of graphs with size range data of the vesicles in the different populations and vesicle concentration data when using NanoSight300.
- Characterization of extracellular vesicles populations by specific markers [ Time Frame: 45 days ]
Analysis of population-specific markers by Western Blot in order to further differentiate endometrial fluid vesicles populations.
The aim is to differentiate populations based on the presence or absence of these markers.
Original Primary Outcome: Same as current
Current Secondary Outcome: Qualitative (names) and quantitative (expression levels) information about the different biomolecules that could be present in the samples: DNA, RNA, proteins, lipids and other small metabolites. [ Time Frame: three years ]
Original Secondary Outcome: Same as current
Information By: Igenomix
Dates:
Date Received: June 2, 2016
Date Started: April 2016
Date Completion: March 2019
Last Updated: June 20, 2016
Last Verified: June 2016
