The Aim of This Study is to Determine the Incidence of Alterations in Fluorescent In Situ Hybridisation (FISH) of Spermatozoa and to Evaluate the Efficacy of the Use of Annexin Columns (MACS®) in Patients With Altered FISH Undergoing In Vitro Fertilisation Techniques

Evaluation of the Efficacy of Annexin Columns (MACS) in Patients With Altered Sperm FISH Undergoing Assisted Reproductive Techniques

250 petitions for FISH of spermatozoa were requested, determining the total and particular incidence in each of the indications collected. The effect of annexin columns was evaluated in 10 couples with altered FISH. In each of the couples, oocytes were pseudo-randomised to the control group and MACS®. All couples received experimental treatment. Mixed logistic regression model were adjusted to evaluate the effect of MACS® on the response variables studied.

Study Overview

• Status: Completed
• Conditions: Infertility, Male
• Intervention / Treatment: Device: Magnetic Activated Cell Sorting

Detailed Description

Prospective pseudo-randomised study carried out in the reproduction unit of the Hospital General Universitario Dr. Balmis in Alicante. The study was divided into two phases, with both being conducted in parallel. The research was approved by the Ethics Committee for Research on Medicines (CEIm) of the Alicante Department of Health. The treatment and use of the clinical data required for this study complied with current legislation, preserving the anonymity of the patients.

Phase I. A total of 250 sperm FISH tests were requested for different indications in men aged 18-55 years with normal karyotype. Of the total number of tests requested, 16 could not be performed due to low sperm count. Finally, 234 samples were analysed, of which 20 had altered sperm FISH.

Phase II. From the previous phase, not all patients with an altered FISH test were included in the second phase of the study, some of them were lost for different reasons (n=10). A total of 10 couples with no previous children in common, negative serology and normal karyotype were included. Females were aged less than 40 years and males less than 50 years at the time of treatment.

The semen sample of each couple was capacitated by density gradient centrifugation (DGC) and swim up, forming the control group. An aliquot was taken that was destined to pass through the annexin columns (MACS® ART, Miltenyl Biotic GmbH, Germany) constituting the study group. The oocytes obtained in each of the couples were pseudo-randomized and inseminated by ICSI with sperm from the study group and the control group. All couples benefited from the experimental treatment since they had the opportunity to inseminate their oocytes with sperm from both groups, thus excluding treatment bias or asymmetry.

To evaluate the efficacy of MACS® on laboratory outcome, fertilization rates, abnormal fertilization, biopsiable embryos and euploid embryos were compared between the study and control group.

• Study Type: Interventional
• Enrollment (Actual): 10
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Spain
  • Alicante, Spain, 03010
    • ISABIAL

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult
• Accepts Healthy Volunteers: No

Description

Inclusion Criteria:

• Women between 18 and 40 years of age. Men between 18 and 55 years of age.
• Couples without any common, previous and healthy children.
• Couples with indication of IVF-ICSI cycle with Preimplantation Genetic Test for Aneuploidy (PGT-A).
• Men with altered sperm FISH results.

Exclusion Criteria:

• Prior voluntary sterilization.
• Existence of documented medical contraindication for infertility treatment.
• Existence of documented medical contraindication to pregnancy.
• Existence of a documented medical situation that seriously interferes with the development of the offspring.
• Impossibility to comply with the treatment for health-related reasons or other family or social environment-related reasons.
• Existence of a documented situation referring to any other circumstance that may seriously interfere with the development of the offspring submitted for consideration to a health care ethics committee or similar body.
• One or both partners with positive serology for HIV, HBV and VDRL.
• One or both partners with partner with altered karyotype.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Triple

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: MACS; Oocytes corresponding to this group were inseminated by spermatozoa selected after the use of MACS.	Device: Magnetic Activated Cell Sorting; The MACS® ART Annexin V System has been designed for the in vitro elimination of apoptotic human spermatozoa in fresh, cryopreserved or otherwise manipulated semen samples. Samples processed by this technique can be subsequently used in any assisted reproduction procedure. Annexin V recognizes externalized phosphatidylserine (ePS) on the plasma membrane of apoptotic spermatozoa. Depletion of the altered sperm begins with magnetic tagging of the sperm. The system selectively retains unwanted sperm by utilizing the natural ability of annexin V to recognize the ePS on the plasma membrane of the altered sperm, because annexin V is conjugated to metal microspheres. The labeled cells pass through a separation column in a fixed magnetic field. Unwanted (altered) spermatozoa are selectively retained in the separation column. Unlabeled spermatozoa pass through the column where they are collected for later use.; Other Names: MACS
No Intervention: Control; Oocytes corresponding to this group were inseminated with spermatozoa obtained after density gradient centrifugation.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Euploid embryo rate	Embryos without chromosomal alterations in relation to the total number of those that were informative.	This variable was taken 2 days after obtaining the genetic result of the embryo biopsy.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Fertilization rate	Number of correctly fertilized zygotes among the total number of mature oocytes inseminated	This variable was taken 17-20 hours after oocyte insemination.
Biopsiable embryo rate	Number of biopsied embryos versus the total number of correctly fertilized zygotes	This variable was taken on the 3rd day of embryonic development, at the time of the embryo biopsy. .

Collaborators and Investigators

• Sponsor: Instituto de Investigación Sanitaria y Biomédica de Alicante

Study record dates

Study Major Dates

• Study Start (Actual): January 1, 2018
• Primary Completion (Actual): March 31, 2022
• Study Completion (Actual): March 31, 2022

Study Registration Dates

• First Submitted: July 21, 2024
• First Submitted That Met QC Criteria: July 25, 2024
• First Posted (Actual): July 29, 2024

Study Record Updates

• Last Update Posted (Actual): July 29, 2024
• Last Update Submitted That Met QC Criteria: July 25, 2024
• Last Verified: July 1, 2024

More Information

Terms related to this study

• Keywords: In Vitro Fertilization; MACS; sperm FISH; annexin columns; Preimplantation Genetic Testing for aneuploidies
• Additional Relevant MeSH Terms: Urogenital Diseases; Male Urogenital Diseases; Genital Diseases, Male; Genital Diseases; Infertility; Infertility, Male
• Other Study ID Numbers: 190767

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No