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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.
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.
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| Label | Type | Description | Intervention Names |
|---|---|---|---|
| men | men undergoing routine semen analysis for infertility |
|
| Name | Type | Description | Arm Group Labels | Other Names |
|---|---|---|---|---|
| cryopreservation | Other | 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 |
| 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 |
| 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 |
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Inclusion Criteria:
Exclusion Criteria:
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Men undergoing routine semen analysis for infertility for reproductive medicine of university hospital in Clermont-Ferrand
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| Name | Affiliation | Role |
|---|---|---|
| Florence BRUGNON | University Hospital, Clermont-Ferrand | Principal Investigator |
| Facility | Status | City | State | ZIP | Country | Contacts |
|---|---|---|---|---|---|---|
| Chu Clermont-Ferrand | Clermont-Ferrand | 63003 | France |
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| ID | Term |
|---|---|
| D015925 | Cryopreservation |
| ID | Term |
|---|---|
| D014021 | Tissue Preservation |
| D016591 | Histocytological Preparation Techniques |
| D003584 | Cytological Techniques |
| D019411 | Clinical Laboratory Techniques |
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|
| 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 |
| D019937 | Diagnostic Techniques and Procedures |
| D003933 | Diagnosis |
| D006652 | Histological Techniques |
| D011309 | Preservation, Biological |
| D013812 | Therapeutics |
| D008919 | Investigative Techniques |