Cryogenic Preservation of Spermatozoa

Study Overview
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Brief Summary
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The reference technique for the conservation of gametes is storage in liquid nitrogen but new vats of nitrogen vapor (storage over liquid nitrogen) or in dry phase (storage in an insulated compartment of liquid nitrogen in a tank Liquid nitrogen) also allow the storage of flakes. The purpose of this work is to evaluate the dry-phase cryopreservation technique of liquid nitrogen compared with liquid-phase storage, depending on the duration of cryopreservation.

Detailed Description
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Objective: To evaluate the effects of cryopreserved sperm in dry and liquid phase nitrogen at 3 and 6 month on sperm numeration, motility, vitality, morphology, acrosomal integrity and DNA fragmentation. Design: Experimental study, investigator was blinded to the type of Cryopreservation. Patient(s): Semen samples were collected from patients who came in laboratory for semen analysis Intervention: Samples were frozen with a programmable freezing unit. Each semen sample was divided into two aliquots. One aliquot was plunged into liquid nitrogen and the other was stored in dry-phase nitrogen for 3 or 6 month. Thawing was performed at room temperature.

Official Title
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Cryogenic Preservation of Spermatozoa : Comparative Study Between Both Dry and Liquid Phase Nitrogen Storages

Conditions
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Fertility, Spermatic Parameters

Intervention / Treatment
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* Other: cryopreservation


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Men undergoing routine semen analysis for infertility for reproductive medicine of university hospital in Clermont-Ferrand Exclusion Criteria: women

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 60 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| men<br>men undergoing routine semen analysis for infertility | Other: cryopreservation<br>* Samples were frozen with a programmable freezing unit. Each semen sample was divided into two aliquots. One aliquot was plunged into liquid nitrogen and the other was stored in dry-phase nitrogen for 3 or 6 month. Thawing was performed at room temperature<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Sperm DNA integrity | the spermatic DNA integrity can be quantified by TUNEL method. The result will be expressed as a percentage of fragmented DNA | at 3 months | 
| Sperm DNA integrity | the spermatic DNA integrity can be quantified by TUNEL method. The result will be expressed as a percentage of fragmented DNA | at 6 months | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Sperm parameter post cryopreservation | The Sperm motility will be measured according to WHO recommendations | at 3 and 6 months | 
| Lab parameter post cryopreservation | The Sperm motility will be measured according to WHO recommendations | at 3 and 6 months | 
| Spermatic DNA Compaction | The spermatic DNA Compaction will be determinated by chromomycine A3 labelling (CMA3). A sperm is good when it has at least 30% of positive spermatozoa to CMA3 assay | at 3 and 6 months | 
| Acrosomal integrity | The acrosomal integrity will be determinated by PSA-FITC labelling. | at 3 and 6 months | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Cryo preservation, Spermatic nuclear quality