Hypotaurine Supplementation Benefits in Cryopreservation

Cryopreservation Clinical Trial

— HYPOTAURINE  

Official title:

Official Title: Hypotaurine Supplementation in Freezing and Preparation Media Improves Human Sperm DeoxyriboNucleic Acid (DNA) and Fertilizing Ability

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT04011813
• Other study ID #: 2014 HYPOTAURINE
• Status: Completed
• Start date: May 1, 2014
• Est. completion date: May 1, 2019

Study information

• Verified date: June 2019
• Source: University Hospital, Clermont-Ferrand
• Contact: n/a
• Is FDA regulated: No
• Study type: Observational

Clinical Trial 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.

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.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 33
• Est. completion date: May 1, 2019
• Est. primary completion date: August 1, 2018
• Accepts healthy volunteers: No
• Gender: Male
• Age group: 18 Years and older

Eligibility

Inclusion Criteria:

• None

Exclusion Criteria:

• None

Related Conditions & MeSH terms

• Cryopreservation
• Human Spermatozoa Parameters
• Hypotaurine Supplementation
• Spermatozoa DNA Alterations

Intervention

Other:
• Hypotaurine : antioxidant and osmoregulator
Hypotaurine has protective effects on sperm motility, capacitation and acrosome reaction and reduces apoptotic markers. Hypotaurine (50mM) was added in density gradient centrifugation, washing and cryopreservation media washing and cryopreservation media before spermatozoa freezing

Locations

Country	Name	City	State
n/a

Sponsors (4)

Lead Sponsor	Collaborator
University Hospital, Clermont-Ferrand	Université d'Auvergne, Centre National de la Recherche Scientifique, Université d'Auvergne, Institut National de la Santéet de la Recherche Médicale, University Hospital, Clermont-Ferrand, Hematology laboratory

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	chromatin packaging labelled using aniline blue and chromomycin A3	Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of chromatin packaging labelled using aniline blue and chromomycin A3	Day 0
Primary	DNA fragmentation using TUNEL assay	Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of DNA fragmentation using TUNEL assay	Day 0
Primary	DNA oxidation assessed by 8-OHdG immunodetections	Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of DNA oxidation assessed by 8-OHdG immunodetections	Day 0
Primary	vacuoles presence in the spermatozoa head using Motile Sperm Organelle Morphology Examination (MSOME)	Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of vacuoles presence in the spermatozoa head using Motile Sperm Organelle Morphology Examination (MSOME)	Day 0
Secondary	vitality using Eosin Nigrosin	Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of vitality using Eosin Nigrosin	Day 0
Secondary	motility total and progressive	Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of motility total and progressive	Day 0
Secondary	integrity of the acrosome using Fluorescein IsoThioCyanate-Pisum Sativum Agglutinin (FITC-PSA) labelling	Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of integrity of the acrosome using Fluorescein IsoThioCyanate-Pisum Sativum Agglutinin (FITC-PSA) labelling	Day 0
Secondary	markers of PKA-dependent capacitation intracellular signaling pathway assessing western blot	Comparison after a cycle of freezing-thawing for both conditions, with or without hypotaurine supplementation of markers of PKA-dependent capacitation intracellular signaling pathway assessing western blot	Day 0