Clinical Trials Logo

Clinical Trial Details — Status: Recruiting

Administrative data

NCT number NCT05383599
Other study ID # 22114CROSP_IM
Secondary ID
Status Recruiting
Phase
First received
Last updated
Start date July 16, 2022
Est. completion date August 1, 2023

Study information

Verified date February 2023
Source Universitair Ziekenhuis Brussel
Contact ileana mateizel
Phone 003224776690
Email ileana.mateizel@uzbrussel.be
Is FDA regulated No
Health authority
Study type Observational

Clinical Trial Summary

The present study aims to determine which of the sperm selection techniques used in IVF (In Vitro Fertilization) laboratory leads to selection of the best/healthiest sperm population based on their morpho-functional parameters. Different procedures are currently used in our laboratory to select the sperm population that will be used to inseminate the oocytes: Density Gradient Centrifugation (DGC), DGC in combination with Magnetic Activated Sperm Sorting (MACS), and Microfluidic Sperm Sorting (MSS). The sperm cells selected by these techniques will be evaluated for specific morpho-functional features such as, motility, morphological abnormalities, and apoptotic status [phosphatidylserine translocation (early stages of apoptosis)] and DNA (Deoxyribonucleic Acid) fragmentation as an indicator of stages of apoptosis.


Description:

Different procedures are currently available to select the sperm population that will be used to inseminate the oocytes during Medical Assisted Reproduction (MAR). The selected population should contain fully functional spermatozoa that possess the ability to reach the fertilization site and allow optimal fertilization and embryo development. These abilities are strictly dependent on specific sperm morpho-functional features such as, but not limited to, motility, morphological abnormalities, apoptotic status [phosphatidylserine translocation (early stages of apoptosis)] and DNA fragmentation (later stages of apoptosis), The present study aims to determine which of the sperm selection techniques used in our IVF laboratory lead to selection of the best/healthy sperm population based on their morpho-functional parameters. For this purpose, unused raw semen after routine diagnostic analysis will be split in 3 fractions and will be processed immediately by different sperm preparation methods: 1) DGC, 2) DGC and MACS, and 3) MSS. The sperm cells selected by each method will be analyzed for morphological and functional parameters. Initial sperm concentration (×106/ml) will be assessed using a counting chamber. Sperm motility will be evaluated on 200 spermatozoa under 40× magnifications and the results will be expressed as percentages. After each selection procedure, concentration and motility will be determined with the use of a Neubauer counting chamber. Spermatozoa with normal morphology will be checked on smears stained using Diff Quick Staining on dried smears, 200 spermatozoa will be classified as normal/abnormal according to Kruger strict criteria. Phosphatidylserine (PS) translocation (early apoptotic marker): will be analyzed using fluorescence microscopy following sperm cell suspension incubation with Annexin V-FITC (Fluorescein Isothiocyanate) and propidium iodide (PI). The spermatozoa will be classified as: viable spermatozoa without PS translocation (PI negative/Annexin negative); viable with translocated PS (PI negative/Annexin positive); dead (PI positive/Annexin positive or PI positive/Annexin negative). DNA fragmentation index (percentage of sperm cells showing DNA fragmentation per number of cells analyzed; DFI) will be analyzed using terminal deoxynucleotidyl-transferase-mediated deoxyuridine triphosphate-fluorescein nick end labeling (TUNEL) assay protocol (commercially available kit "In Situ Cell Death Detection"; Roche Diagnostic). The cleavage of genomic DNA during apoptosis leads to both single-strand breaks (nicks) and double-stranded, low-molecular-weight DNA fragments. These DNA strand breaks can be identified in an enzymatic reaction in two stages: (1) labeling of DNA strand breaks with TdT (Terminal Deoxynucleotidyl Transferase), and (2) incorporation of Fluorescein isothiocyanate (FITC)-dUTP (deoxyuridine triphosphate) into nucleotide polymers. TUNEL-positive sperm stain green and TUNEL-negative samples stain red and it can be directly detected and quantified by fluorescence microscopy. This kit is designed to be a precise, fast, and simple nonradioactive technique to detect and quantify the number apoptotic cells. It is specific as it labels DNA strand breaks generated during apoptosis, which enables the test to discriminate between apoptotic and necrotic cells. Descriptive statistics will be used to report mean, median and minimum and maxi-mum values for each variable. Exact test for Friedman's test will be used to investigate the presence of differences across the 3 techniques. Exact test for Wilcoxon signed rank test will be used for pairwise comparisons. A Bonferroni correction will be applied to adjust for multiple comparisons. Level of significance is set at p<0.05, 2-tailed. Statistical analysis will be performed by means of STATA 15.1 and SPSS 26.0 (Statistical Package for the Social Sciences). A number of 50 patients will be included in the study. The results obtained will help to improve the sperm selection process in the IVF laboratory


Recruitment information / eligibility

Status Recruiting
Enrollment 50
Est. completion date August 1, 2023
Est. primary completion date January 16, 2023
Accepts healthy volunteers
Gender Male
Age group N/A and older
Eligibility Inclusion Criteria: - semen volume =1.5ml - sperm concentration >20 x 106 spermatozoa/ml - =30% progressive motility Exclusion Criteria: - semen samples processed for immediate or later use in Assisted Reproductive Medicine

Study Design


Related Conditions & MeSH terms


Locations

Country Name City State
Belgium UZ Brussel CRG Brussels

Sponsors (1)

Lead Sponsor Collaborator
Universitair Ziekenhuis Brussel

Country where clinical trial is conducted

Belgium, 

Outcome

Type Measure Description Time frame Safety issue
Primary DNA Fragmentation Index percentage of sperm cells with DNA fragmentation out of selected sperm cells 1 year
Primary Progressive Motility Index percentage of sperm cells with progressive motility out of selected sperm cells 1 year
Primary Early Apoptosis Index percentage of early apoptotic sperm cells out of selected sperm cells 1 year
Primary Normal Morphology Index percentage of morphological normal sperm cells out of selected sperm cells 1 year
Primary Concentration number of sperm cells per ml of ejaculated semen sample 1 year
See also
  Status Clinical Trial Phase
Recruiting NCT05503862 - Home Semen Testing in Men Beginning Attempts to Conceive N/A
Recruiting NCT03307655 - Relationship Between Nitric Oxide (NO) in Follicular Fluid and Sperm Fertilization Ability N/A
Withdrawn NCT02839447 - A Clinical Evaluation of Semen Quality Using the MiOXSYS™ System 2.0
Completed NCT00975117 - Spermotrend in the Treatment of Male Infertility Phase 3
Completed NCT00756561 - HOP-2A - Intratesticular Hormone Levels N/A
Recruiting NCT04541459 - Validation of New Devices Against Ambient Electromagnetic Radiation Early Phase 1
Completed NCT02889341 - Valuation of Variable Dose of Docosahexanoic Acid for the Improvement of the Parameters of Male Fertility N/A
Not yet recruiting NCT03167008 - Vitamin D Level vs Semen Parameters
Completed NCT02268123 - Correlating In Vitro Fertilization Outcomes After Euploid Blastocyst Transfer With Sperm DNA Fragmentation
Completed NCT01232465 - Impact of Sperm DNA Integrity on In Vitro Cycles N/A
Recruiting NCT00119925 - 'SPRING'-Study: "Subfertility Guidelines: Patient Related Implementation in the Netherlands Among Gynaecologists" N/A
Completed NCT03552081 - Tobacco and Sperm Genome: Effects of Smoking Cessation N/A
Recruiting NCT05200663 - Efficacy Comparison of Tamoxifen and Tamoxifen With Antioxidants on Semen Quality of Male With Idiopathic Infertility Phase 2
Completed NCT02025270 - MSCs For Treatment of Azoospermic Patients Phase 1/Phase 2
Recruiting NCT06147700 - Identifying Molecular Determinants of Infertility in Men (MODIFY)
Recruiting NCT06188936 - Home Semen Analysis Tests as a Screening Tool for Fertility Patients N/A
Recruiting NCT04200118 - Epigenetic and Genetic Effects in Cancer Patients: Analysis Pre and After Treatment
Terminated NCT02605070 - Pilot Study on the Effects of FSH Treatment on the Epigenetic Characteristics of Spermatozoa in Infertile Patients With Severe Oligozoospermia Phase 3
Completed NCT01407432 - Impact of Folates in the Care of the Male Infertility Phase 3
Completed NCT00596739 - A Study of the Pre- and Post-operative Semen Analyses and Reproductive Hormone Levels of Men Undergoing Weight-reduction Surgery N/A