Clinical trial NCT04011813: Hypotaurine Supplementation Benefits in Cryopreservation
Official title: Official Title: Hypotaurine Supplementation in Freezing and Preparation Media Improves Human Sperm DeoxyriboNucleic Acid (DNA) and Fertilizing Ability
Study type: OBSERVATIONAL
Overall status: COMPLETED
Conditions: Cryopreservation
Human Spermatozoa Parameters
Hypotaurine Supplementation
Spermatozoa DNA Alterations
Keywords: cryopreservation
density gradient centrifugation
DNA fragmentation, oxidation, chromatin packaging
antioxidant hypotaurine supplementation
cryo-capacitation
human spermatozoa fertilizing ability
Lead sponsor: University Hospital, Clermont-Ferrand
Sponsor class: OTHER
Collaborators: name: Université d'Auvergne, Institut National de la Santéet de la Recherche Médicale
name: Université d'Auvergne, Centre National de la Recherche Scientifique
name: University Hospital, Clermont-Ferrand, Hematology laboratory
First posted: 2019-07-09
Enrollment: 33
Sex: MALE
Minimum age (years): 18
Standard ages: ADULT
OLDER_ADULT
Accepts healthy volunteers: no
Observational model: COHORT
Time perspective: PROSPECTIVE
Study population: Men undergoing routine semen analysis at the Center for Reproductive Medicine

Brief summary: Although it is widely used, slow freezing can induce strong functional and nuclear spermatic alterations reducing the chances of pregnancy. The study objective is to determinate the effects of the combination of hypotaurine supplementation and spermatozoa selection by Density Gradient Centrifugation (DGC) on human sperm functions and DNA quality during a freezing-thawing cycle.

Detailed description: This prospective study was performed on surplus semen after a density gradient centrifugation-frozen-thawing cycle. Samples were obtained from men undergoing routine semen analysis at the Center for Reproductive Medicine. Spermatozoa were selected by density gradient centrifugation, washed and frozen using a programmable device. Each step was performed in parallel with (H+ arm) or without (H- arm) 50mM hypotaurine supplementation. After thawing, investigator team compared for both conditions the total and progressive mobility, vitality, integrity of the acrosome, markers of Protein Kinase A (PKA) dependent capacitation intracellular signaling pathway and nuclear quality by measuring chromatin packaging, DNA fragmentation and oxidation and vacuoles presence in the spermatozoa head.

Eligibility criteria: Inclusion Criteria:

* None

Exclusion Criteria:

* None

Primary outcomes:
measure: chromatin packaging labelled using aniline blue and chromomycin A3
timeFrame: Day 0
measure: DNA fragmentation using TUNEL assay
timeFrame: Day 0
measure: DNA oxidation assessed by 8-OHdG immunodetections
timeFrame: Day 0
measure: vacuoles presence in the spermatozoa head using Motile Sperm Organelle Morphology Examination (MSOME)
timeFrame: Day 0
Secondary outcomes:
measure: vitality using Eosin Nigrosin
timeFrame: Day 0
measure: motility total and progressive
timeFrame: Day 0
measure: integrity of the acrosome using Fluorescein IsoThioCyanate-Pisum Sativum Agglutinin (FITC-PSA) labelling
timeFrame: Day 0
measure: markers of PKA-dependent capacitation intracellular signaling pathway assessing western blot
timeFrame: Day 0
Other outcomes:
Locations:
Study officials:
affiliation: University Hospital, Clermont-Ferrand
name: Florence BRUGNON, MD, PhD
role: PRINCIPAL_INVESTIGATOR
References:
citation: Pons-Rejraji H, Vorilhon S, Difrane A, Dollet S, Bourgne C, Berger M, Chaput L, Pereira B, Bouche C, Drevet JR, Brugnon F. Beneficial effects of hypotaurine supplementation in preparation and freezing media on human sperm cryo-capacitation and DNA quality. Basic Clin Androl. 2021 Nov 4;31(1):26. doi: 10.1186/s12610-021-00144-6.