Human Sperm Markers of Cryodamage Resistance
Study Details
Study Description
Brief Summary
Sperm freezing has been employed for decades for male fertility preservation in cases of foreseeable or unexpected loss of fertility to guarantee future paternity, and also as a complement of assisted reproduction techniques.
Sperm quality after thawing is highly variable, even among consecutive samples from the same individual, with mean survival rates around 40%. To date, the molecular basis of the adequate resistance or intolerance to freezing/thawing protocols is unknown, and its knowledge can lead to improvement in the selection of the samples to be frozen and also in the adequate supplement of cryopreservation media. Microarray analysis provides a powerful tool to address the molecular explanation beyond this behaviour, yielding results about comparative messenger ribonucleic acid (mRNA)expression under the two different biological conditions: optimal and suboptimal survival.
Then, the aim of the investigators' study is to determine the genomic profile of sperm samples depending on their survival resistance to cryopreservation, to determine genes involved in cryodamage sensitivity.
| Condition or Disease | Intervention/Treatment | Phase |
|---|---|---|
|
Detailed Description
Nested cases and controls design, with donor sperm samples frozen under conventional protocols, categorized depending on their good (GSR, n=20) (less than 20% motility decrease) or bad (BSR, n=20) (more than 20% motility decrease) survival rates. Sperm mRNA was extracted using Trizol protocol, suspended in diethylpyrocarbonate (DEPC)-treated water and frozen at -80 degrees until the microarray experiments were performed. RNAs were analyzed on Agilent Bioanalyzer 2100. Samples were pooled in 4 (5 samples/pool and 4 pools/group). Finally, 8 Agilent One colour Whole genome microarray (44K) were performed with pooled samples, 4 microarrays per group.
The results will be evaluate to detect those genes differentially expressed.
Study Design
Arms and Interventions
| Arm | Intervention/Treatment |
|---|---|
| High cryoresistance High resistance to sperm cryopreservation (decreased sperm motility <20%) |
Other: Sperm cryopreservation
Sperm from the different groups are cryopreserved to evaluate their resistance to preservation.
|
| Low cryoresistance Low resistance to sperm cryopreservation (decreased sperm motility >20%) |
Other: Sperm cryopreservation
Sperm from the different groups are cryopreserved to evaluate their resistance to preservation.
|
Outcome Measures
Primary Outcome Measures
- Genomic profile of sperm samples depending on their survival resistance to cryopreservation [A year and a half]
The genomic profile of sperm samples depending on their survival resistance to cryopreservation will be analysed and expressed as: Common or differentially expressed genes: in case of genes expressed in both groups Exclusive Genes, in case of genes expressed just in one group
Secondary Outcome Measures
- Pregnancy rates after artificial insemination with donor semen in the different groups. [two years]
Once obtained the genomic profile of samples from donors included the different groups, we will analyze its relationship with the reproductive outcomes, in particular pregnancy rates after conventional artificial insemination (percentage of positive pregnancy test)
Eligibility Criteria
Criteria
Inclusion Criteria:
The criteria used for the inclusion of donors in the sperm bank will be considered:
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males from 18 to 35 years.
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good physical and psychological conditions and no hereditary diseases
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seminal parameters: concentration > 40 million sperm / ml; percentage of progressive motility> 50% percentage of normal forms> 4%.
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For pregnancy rates after artificial insemination, only women< 40 years without tubal pathologies and artificial insemination indications will be included.
Exclusion Criteria:
All the population not included in the list of inclusion will be excluded.
Contacts and Locations
Locations
| Site | City | State | Country | Postal Code | |
|---|---|---|---|---|---|
| 1 | IVI Alicante | Alicante | Spain | 03015 |
Sponsors and Collaborators
- Instituto Valenciano de Infertilidad, IVI Alicante
- Igenomix
Investigators
- Principal Investigator: Elena Sellés, PhD, Instituto Valenciano de Infertilidad, IVI Alicante
Study Documents (Full-Text)
None provided.More Information
Publications
- Casas I, Sancho S, Briz M, Pinart E, Bussalleu E, Yeste M, Bonet S. Freezability prediction of boar ejaculates assessed by functional sperm parameters and sperm proteins. Theriogenology. 2009 Oct 15;72(7):930-48. doi: 10.1016/j.theriogenology.2009.07.001. Epub 2009 Aug 3.
- Gadea J, Molla M, Selles E, Marco MA, Garcia-Vazquez FA, Gardon JC. Reduced glutathione content in human sperm is decreased after cryopreservation: Effect of the addition of reduced glutathione to the freezing and thawing extenders. Cryobiology. 2011 Feb;62(1):40-6. doi: 10.1016/j.cryobiol.2010.12.001. Epub 2010 Dec 13.
- García-Herrero S, Garrido N, Martínez-Conejero JA, Remohí J, Pellicer A, Meseguer M. Ontological evaluation of transcriptional differences between sperm of infertile males and fertile donors using microarray analysis. J Assist Reprod Genet. 2010 Feb;27(2-3):111-20. doi: 10.1007/s10815-010-9388-5. Epub 2010 Feb 2.
- García-Herrero S, Meseguer M, Martínez-Conejero JA, Remohí J, Pellicer A, Garrido N. The transcriptome of spermatozoa used in homologous intrauterine insemination varies considerably between samples that achieve pregnancy and those that do not. Fertil Steril. 2010 Sep;94(4):1360-1373. doi: 10.1016/j.fertnstert.2009.07.1671. Epub 2009 Sep 30.
- Garrido N, Martínez-Conejero JA, Jauregui J, Horcajadas JA, Simón C, Remohí J, Meseguer M. Microarray analysis in sperm from fertile and infertile men without basic sperm analysis abnormalities reveals a significantly different transcriptome. Fertil Steril. 2009 Apr;91(4 Suppl):1307-10. doi: 10.1016/j.fertnstert.2008.01.078. Epub 2008 Mar 25.
- Holt WV, Medrano A, Thurston LM, Watson PF. The significance of cooling rates and animal variability for boar sperm cryopreservation: insights from the cryomicroscope. Theriogenology. 2005 Jan 15;63(2):370-82. Review.
- Rahana AR, Ng SP, Leong CF, Rahimah MD. Comparison between mechanical freezer and conventional freezing using liquid nitrogen in normozoospermia. Singapore Med J. 2011 Oct;52(10):734-7.
- Thurston LM, Siggins K, Mileham AJ, Watson PF, Holt WV. Identification of amplified restriction fragment length polymorphism markers linked to genes controlling boar sperm viability following cryopreservation. Biol Reprod. 2002 Mar;66(3):545-54.
- Thurston LM, Watson PF, Holt WV. Semen cryopreservation: a genetic explanation for species and individual variation? Cryo Letters. 2002 Jul-Aug;23(4):255-62. Review.
- 1406-ALC-048-ES