Optimization of the Isolation Protocol for Extracellular Vesicles (EVs) From Human Follicular Fluid

NCT ID: NCT06952751

Last Updated: 2025-08-15

Study Results

Results pending

The study team has not published outcome measurements, participant flow, or safety data for this trial yet. Check back later for updates.

Basic Information

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Recruitment Status

RECRUITING

Clinical Phase

NA

Total Enrollment

4 participants

Study Classification

INTERVENTIONAL

Study Start Date

2025-07-20

Study Completion Date

2025-12-31

Brief Summary

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Background: Extracellular vesicles (EVs) are membrane-bound vesicles found in all biological fluids, containing various regulatory molecules, including microRNAs (miRNAs). It has been suggested that the EVs in human follicular fluid (the fluid surrounding the oocyte within the ovary) play a crucial role in oocyte development through these miRNAs. However, several methods exist for isolating these EVs from follicular fluid. Before further research can be conducted on the pathophysiology and potential diagnostic and therapeutic applications of these EVs and their content, the optimal isolation technique must be determined for future studies.

Each method has its advantages and disadvantages, and the purity and efficiency may vary between species and different biological fluids. There are only a limited number of studies comparing the various techniques for application in human follicular fluid.

Objective: This study will compare the three most commonly used techniques for isolating EVs: size-based, sedimentation-based (differential ultracentrifugation), and buoyancy-based isolation techniques.

Methodology:

In this prospective study, follicular fluid will be collected from four patients undergoing transvaginal oocyte retrieval following ovarian stimulation at Ghent University Hospital. Patients over 40 years of age or with endometriosis, adenomyosis, or PCOS (polycystic ovarian syndrome) will not be included in this study. During the oocyte retrieval procedure, follicular fluid will be collected from 2-4 oocytes per patient. Since follicular fluid is not used in the patient's treatment (residual material), it does not affect the treatment.

After collection, the three isolation techniques will be applied to each sample. The techniques will be compared in terms of purity (absence of contamination) and efficiency (EV concentration) using transmission electron microscopy, nanoparticle tracking analysis, and western blotting.

Detailed Description

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Conditions

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Extracellular Vesicles

Study Design

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Allocation Method

NA

Intervention Model

SINGLE_GROUP

All collections of follicular fluid will be divided to test all three isolation methods under investigation.
Primary Study Purpose

BASIC_SCIENCE

Blinding Strategy

NONE

Study Groups

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Intervention group

Intervention group

Group Type EXPERIMENTAL

Differential ultracentrifugation

Intervention Type OTHER

Differential ultracentrifugation

OptiPrep™ density gradient ultracentrifugation

Intervention Type OTHER

OptiPrep™ density gradient ultracentrifugation

Size-exclusion chromatography

Intervention Type OTHER

Size-exclusion chromatography

Combined techniques

Intervention Type OTHER

Combined techniques

Interventions

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Differential ultracentrifugation

Differential ultracentrifugation

Intervention Type OTHER

OptiPrep™ density gradient ultracentrifugation

OptiPrep™ density gradient ultracentrifugation

Intervention Type OTHER

Size-exclusion chromatography

Size-exclusion chromatography

Intervention Type OTHER

Combined techniques

Combined techniques

Intervention Type OTHER

Eligibility Criteria

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Inclusion Criteria

* Patients planned for an oocyte retrieval at Ghent University Hospital.

Exclusion Criteria

* Diagnosis of endometriosis/adenomyosis
* Diagnosis of PCOS (Polycystic Ovary Syndrome)
* Macroscopic blood contamination at oocyte retrieval
Minimum Eligible Age

18 Years

Maximum Eligible Age

40 Years

Eligible Sex

FEMALE

Accepts Healthy Volunteers

No

Sponsors

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University Ghent

OTHER

Sponsor Role collaborator

University Hospital, Ghent

OTHER

Sponsor Role lead

Responsible Party

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Responsibility Role SPONSOR

Locations

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Ghent University Hospital

Ghent, , Belgium

Site Status RECRUITING

Countries

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Belgium

Central Contacts

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Dominic Stoop, Prof. dr.

Role: CONTACT

+ 329 332 34 42

Eva Decroos, Dr.

Role: CONTACT

+329 332 67 56

Facility Contacts

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Eva Decroos, MD

Role: primary

+3293326756

Kathleen Wijnant, MSc

Role: backup

+3293323442

Other Identifiers

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G096624N

Identifier Type: OTHER_GRANT

Identifier Source: secondary_id

ONZ-2024-0419

Identifier Type: -

Identifier Source: org_study_id

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