Molecular Diagnostic of Sperm Cryoresistance of Semen Donors and Clinical Outcomes After Artificial Insemination. Optimization of Sperm Bank
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 发起方
- 入组人数
- 40
- 试验地点
- 1
- 主要终点
- Genomic profile of sperm samples depending on their survival resistance to cryopreservation
研究概览
简要总结
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.
详细描述
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.
研究设计
- 研究类型
- Observational
- 观察模型
- Cohort
- 时间视角
- Prospective
入排标准
- 年龄范围
- 18 Years 至 35 Years（Adult）
- 性别
- Male
- 接受健康志愿者
- 是
入选标准
- •The criteria used for the inclusion of donors in the sperm bank will be considered:
- •males from 18 to 35 years.
- •good physical and psychological conditions and no hereditary diseases
- •seminal parameters: concentration > 40 million sperm / ml; percentage of progressive motility> 50% percentage of normal forms> 4%.
- •For pregnancy rates after artificial insemination, only women< 40 years without tubal pathologies and artificial insemination indications will be included.
排除标准
- •All the population not included in the list of inclusion will be excluded.
结局指标
主要结局
Genomic profile of sperm samples depending on their survival resistance to cryopreservation
时间窗: 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
次要结局
- Pregnancy rates after artificial insemination with donor semen in the different groups.(two years)
研究者
Elena Sellés Soriano
Molecular diagnostic of sperm cryoresistance of semen donors and clinical outcomes after artificial insemination. Optimization of sperm bank
Instituto Valenciano de Infertilidad, IVI Alicante