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

NCT06952751 | Recruiting

• 2 other identifiers: ONZ-2024-0419G096624N
• Study Type: interventional
• Target: 4
• Locations: 1 country
• Sites: 1

Brief Summary

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.

Conditions

Extracellular Vesicles

Keywords

Extracellular vesicles; Isolation

Outcome Measures

Primary Outcomes (1)

• Purety

  Purety of EV isolation

  3 months

Secondary Outcomes (2)

• Efficiency

  3 months

• RNA efficiency

  6 months

Study Arms (1)

Intervention group

EXPERIMENTAL

Intervention group

Other: Differential ultracentrifugation; Other: OptiPrep™ density gradient ultracentrifugation; Other: Size-exclusion chromatography; Other: Combined techniques

Interventions

Differential ultracentrifugation OTHER

Differential ultracentrifugation

Intervention group

OptiPrep™ density gradient ultracentrifugation OTHER

OptiPrep™ density gradient ultracentrifugation

Intervention group

Size-exclusion chromatography OTHER

Size-exclusion chromatography

Intervention group

Combined techniques OTHER

Combined techniques

Intervention group

Eligibility Criteria

• Age: 18 Years - 40 Years
• Sex: female
• Healthy Volunteers: No
• Age Groups: Adult (18-64)

You may qualify if:

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

You may not qualify if:

• Diagnosis of endometriosis/adenomyosis
• Diagnosis of PCOS (Polycystic Ovary Syndrome)
• Macroscopic blood contamination at oocyte retrieval

Sponsors & Collaborators

• University Hospital, Ghent: lead
• University Ghent: collaborator

Study Sites (1)

Ghent University Hospital

Ghent, 9000, Belgium

RECRUITING

MeSH Terms

Interventions

Chromatography, Gel

Intervention Hierarchy (Ancestors)

Chromatography, Liquid; Chromatography; Chemistry Techniques, Analytical; Investigative Techniques

Central Study Contacts

Dominic Stoop, Prof. dr.

CONTACT

+ 329 332 34 42; arg.studies@uzgent.be

Eva Decroos, Dr.

CONTACT

+329 332 67 56; arg.studies@uzgent.be

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NA
• Masking: NONE
• Purpose: BASIC SCIENCE
• Intervention Model: SINGLE GROUP
• Sponsor Type: OTHER
• Responsible Party: SPONSOR

Model Details: All collections of follicular fluid will be divided to test all three isolation methods under investigation.

Study Record Dates

• First Submitted: April 23, 2025
• First Posted: May 1, 2025
• Study Start: July 20, 2025
• Primary Completion: September 1, 2025
• Study Completion: December 1, 2025
• Last Updated: August 15, 2025

Record last verified: 2025-08

Data Sharing

• IPD Sharing: Will not share

Locations

BE (1)