Human Sperm Cryopreservation as an Alternative to the Decline of Sperm Quality With Aging
8 de septiembre de 2026 actualizado por: IVI Murcia
This observational study will evaluate whether sperm cryopreservation can help preserve sperm quality and molecular characteristics that may be affected by increasing paternal age.
Men aged 18 years and older will provide a semen sample and a blood sample for laboratory analysis.
Each semen sample will be divided into fresh, slow-frozen, and vitrified aliquots.
The study will compare sperm quality (as assessed by computer-assisted sperm analysis), DNA methylation profiles, DNA fragmentation, sperm telomere length before and after cryopreservation.
The effects of age on these parameters will also be assessed.
The impact of age on seminal plasma extracellular vesicle composition and blood cell telomere length will also be analysed.
The goal is to determine whether sperm cryopreservation may represent a strategy for preserving sperm characteristics associated with younger paternal age.
Descripción general del estudio
Estado
Reclutamiento
Descripción detallada
The aging process is associated with declining fertility and increased risks of adverse reproductive and offspring health outcomes. Although the effects of maternal age have been extensively studied, the impact of paternal aging on sperm quality, reproductive outcomes, and molecular characteristics of sperm remains incompletely understood.
The primary objective of this study is to evaluate whether sperm cryopreservation may serve as an alternative to the use of sperm collected at older ages in assisted reproduction treatments. The primary endpoint is the difference in sperm DNA methylation patterns between fresh ejaculated samples and samples subjected to cryopreservation. Cryopreservation will be considered a potentially valid strategy if DNA methylation patterns remain substantially preserved following the cryopreservation process.
At least 45 male participants will be recruited and grouped according to age. Participants will provide one semen sample and a peripheral blood sample. Following routine semen analysis, each semen sample will be divided into three aliquots: fresh, slow-frozen, and vitrified. Cryopreserved aliquots will be stored in liquid nitrogen and subsequently thawed or warmed for analysis.
Standard semen parameters, including volume, concentration, motility, sperm kinetics, and morphology, will be evaluated. Additional analyses will include sperm DNA methylation, DNA fragmentation, telomere length, and characterization of extracellular vesicles and their composition. Identical assessments will be performed on fresh and cryopreserved samples to compare the effects of slow freezing and vitrification.
Secondary objectives include evaluating age-related differences in semen quality and molecular biomarkers, identifying markers associated with biological gamete age, characterizing age-related differences in seminal extracellular vesicles, and exploring associations between sperm molecular characteristics and lifestyle. Participants will also complete validated questionnaires regarding lifestyle and dietary habits.
Tipo de estudio
De observación
Inscripción (Estimado)
45
Contactos y Ubicaciones
Esta sección proporciona los datos de contacto de quienes realizan el estudio e información sobre dónde se lleva a cabo este estudio.
Ubicaciones de estudio
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Murcia
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Murcia, Murcia, España, 30007
- Reclutamiento
- IVI Murcia
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Contacto:
- Juan Carlos Martínez
- Número de teléfono: +34 651 79 10 00
- Correo electrónico: JuanCarlos.Martinez@ivirma.com
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Criterios de participación
Los investigadores buscan personas que se ajusten a una determinada descripción, denominada criterio de elegibilidad. Algunos ejemplos de estos criterios son el estado de salud general de una persona o tratamientos previos.
Criterio de elegibilidad
Edades elegibles para estudiar
- Adulto
- Adulto Mayor
Acepta Voluntarios Saludables
Sí
Método de muestreo
Muestra no probabilística
Población de estudio
Adult men aged 18-65 years attending IVI Murcia for semen analysis, fertility assessment, semen donation, or assisted reproduction treatment who meet the study eligibility criteria and provide informed consent.
Participants will provide semen and blood samples for laboratory assessment of sperm quality and molecular characteristics before and after cryopreservation.
Descripción
Inclusion Criteria:
- Male participants aged 18 to 65 years
- Attending IVI Murcia for semen analysis and/or assisted reproduction treatment
- Able and willing to provide written informed consent
- Willing to provide a semen sample and a blood sample for research purposes
Exclusion Criteria:
- Previous diagnosis of azoospermia or severe oligozoospermia (sperm concentration <1 million sperm/mL)
- Known chromosomal abnormality (abnormal karyotype)
- Known Y chromosome microdeletion
- Febrile illness within the previous 3 months
- Presence of varicocele
- Current use of vitamin or antioxidant supplements intended to improve semen quality
Plan de estudios
Esta sección proporciona detalles del plan de estudio, incluido cómo está diseñado el estudio y qué mide el estudio.
¿Cómo está diseñado el estudio?
Detalles de diseño
Cohortes e Intervenciones
Grupo / Cohorte |
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Men aged 18-39 years
Male participants aged 18-39 years who provide semen and blood samples for laboratory assessment of sperm quality and molecular characteristics.
Semen samples are divided into fresh, slow-frozen, and vitrified aliquots for comparative analysis.
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Men aged 40-49 years
Male participants aged 40-49 years who provide semen and blood samples for laboratory assessment of sperm quality and molecular characteristics.
Semen samples are divided into fresh, slow-frozen, and vitrified aliquots for comparative analysis.
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Men aged ≥50 years
Male participants aged 50 years and older who provide semen and blood samples for laboratory assessment of sperm quality and molecular characteristics.
Semen samples are divided into fresh, slow-frozen, and vitrified aliquots for comparative analysis.
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¿Qué mide el estudio?
Medidas de resultado primarias
Medida de resultado |
Medida Descripción |
Periodo de tiempo |
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Sperm DNA methylation profile
Periodo de tiempo: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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DNA methylation levels of a predefined nine-gene panel measured by pyrosequencing with duplicate measurements and compared between fresh, slow-frozen, and vitrified sperm samples.
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Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Medidas de resultado secundarias
Medida de resultado |
Medida Descripción |
Periodo de tiempo |
|---|---|---|
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Sperm concentration
Periodo de tiempo: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Sperm concentration (million sperm/mL) measured using computer-assisted sperm analysis (CASA).
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Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Total sperm motility
Periodo de tiempo: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Percentage of motile sperm measured using CASA.
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Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Progressive sperm motility
Periodo de tiempo: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Percentage of progressively motile sperm measured using computer-assisted sperm analysis (CASA).
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Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Normal sperm morphology
Periodo de tiempo: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Percentage of sperm with normal morphology assessed by microscopic evaluation of stained slides according to World Health Organization (WHO) criteria.
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Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Sperm DNA fragmentation
Periodo de tiempo: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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DNA fragmentation measured in fresh, slow-frozen, and vitrified sperm samples.
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Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Sperm telomere length
Periodo de tiempo: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Sperm telomere length measured in fresh, slow-frozen, and vitrified sperm samples.
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Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
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Seminal plasma extracellular vesicle composition
Periodo de tiempo: Baseline (study semen collection visit).
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Characterization of seminal plasma extracellular vesicle composition in fresh semen samples.
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Baseline (study semen collection visit).
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Otras medidas de resultado
Medida de resultado |
Medida Descripción |
Periodo de tiempo |
|---|---|---|
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Blood leukocyte telomere length
Periodo de tiempo: Baseline (study blood collection visit).
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Blood cell telomere length measured to investigate its relationship with participant age and sperm molecular characteristics.
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Baseline (study blood collection visit).
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Colaboradores e Investigadores
Aquí es donde encontrará personas y organizaciones involucradas en este estudio.
Patrocinador
Colaboradores
Publicaciones y enlaces útiles
La persona responsable de ingresar información sobre el estudio proporciona voluntariamente estas publicaciones. Estos pueden ser sobre cualquier cosa relacionada con el estudio.
Publicaciones Generales
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- Hezavehei M, Sharafi M, Kouchesfahani HM, Henkel R, Agarwal A, Esmaeili V, Shahverdi A. Sperm cryopreservation: A review on current molecular cryobiology and advanced approaches. Reprod Biomed Online. 2018 Sep;37(3):327-339. doi: 10.1016/j.rbmo.2018.05.012. Epub 2018 Aug 22.
- Wang W, Todorov P, Pei C, Wang M, Isachenko E, Rahimi G, Mallmann P, Isachenko V. Epigenetic Alterations in Cryopreserved Human Spermatozoa: Suspected Potential Functional Defects. Cells. 2022 Jul 4;11(13):2110. doi: 10.3390/cells11132110.
- Wang D, Jueraitetibaike K, Tang T, Wang Y, Jing J, Xue T, Ma J, Cao S, Lin Y, Li X, Ma R, Chen X, Yao B. Seminal Plasma and Seminal Plasma Exosomes of Aged Male Mice Affect Early Embryo Implantation via Immunomodulation. Front Immunol. 2021 Oct 12;12:723409. doi: 10.3389/fimmu.2021.723409. eCollection 2021.
- Tao Y, Sanger E, Saewu A, Leveille MC. Human sperm vitrification: the state of the art. Reprod Biol Endocrinol. 2020 Mar 7;18(1):17. doi: 10.1186/s12958-020-00580-5.
- Sysoeva AP, Makarova NP, Silachev DN, Lobanova NN, Shevtsova YA, Bragina EE, Kalinina EA, Sukhikh GT. Influence of Extracellular Vesicles of the Follicular Fluid on Morphofunctional Characteristics of Human Sperm. Bull Exp Biol Med. 2021 Dec;172(2):254-262. doi: 10.1007/s10517-021-05372-4. Epub 2021 Dec 2.
- Khosravizadeh Z, Khodamoradi K, Rashidi Z, Jahromi M, Shiri E, Salehi E, Talebi A. Sperm cryopreservation and DNA methylation: possible implications for ART success and the health of offspring. J Assist Reprod Genet. 2022 Aug;39(8):1815-1824. doi: 10.1007/s10815-022-02545-6. Epub 2022 Jun 17.
- Sciamanna I, Serafino A, Shapiro JA, Spadafora C. The active role of spermatozoa in transgenerational inheritance. Proc Biol Sci. 2019 Aug 28;286(1909):20191263. doi: 10.1098/rspb.2019.1263. Epub 2019 Aug 28.
- Reshi QUA, Godakumara K, Ord J, Dissanayake K, Hasan MM, Andronowska A, Heath P, Fazeli A. Spermatozoa, acts as an external cue and alters the cargo and production of the extracellular vesicles derived from oviductal epithelial cells in vitro. J Cell Commun Signal. 2023 Sep;17(3):737-755. doi: 10.1007/s12079-022-00715-w. Epub 2022 Dec 5.
- Practice Committee of the American Society for Reproductive Medicine. The clinical utility of sperm DNA integrity testing: a guideline. Fertil Steril. 2013 Mar 1;99(3):673-7. doi: 10.1016/j.fertnstert.2012.12.049. Epub 2013 Feb 1.
- Njajou OT, Cawthon RM, Damcott CM, Wu SH, Ott S, Garant MJ, Blackburn EH, Mitchell BD, Shuldiner AR, Hsueh WC. Telomere length is paternally inherited and is associated with parental lifespan. Proc Natl Acad Sci U S A. 2007 Jul 17;104(29):12135-9. doi: 10.1073/pnas.0702703104. Epub 2007 Jul 10.
- Machtinger R, Laurent LC, Baccarelli AA. Extracellular vesicles: roles in gamete maturation, fertilization and embryo implantation. Hum Reprod Update. 2016 Mar-Apr;22(2):182-93. doi: 10.1093/humupd/dmv055. Epub 2015 Dec 9.
- Lismer A, Kimmins S. Emerging evidence that the mammalian sperm epigenome serves as a template for embryo development. Nat Commun. 2023 Apr 14;14(1):2142. doi: 10.1038/s41467-023-37820-2.
- Laurentino S, Cremers JF, Horsthemke B, Tuttelmann F, Czeloth K, Zitzmann M, Pohl E, Rahmann S, Schroder C, Berres S, Redmann K, Krallmann C, Schlatt S, Kliesch S, Gromoll J. A germ cell-specific ageing pattern in otherwise healthy men. Aging Cell. 2020 Oct;19(10):e13242. doi: 10.1111/acel.13242. Epub 2020 Sep 20.
- Kumar P, Wang M, Isachenko E, Rahimi G, Mallmann P, Wang W, von Brandenstein M, Isachenko V. Unraveling Subcellular and Ultrastructural Changes During Vitrification of Human Spermatozoa: Effect of a Mitochondria-Targeted Antioxidant and a Permeable Cryoprotectant. Front Cell Dev Biol. 2021 Jul 2;9:672862. doi: 10.3389/fcell.2021.672862. eCollection 2021.
- Klaver R, Bleiziffer A, Redmann K, Mallidis C, Kliesch S, Gromoll J. Routine cryopreservation of spermatozoa is safe--evidence from the DNA methylation pattern of nine spermatozoa genes. J Assist Reprod Genet. 2012 Sep;29(9):943-50. doi: 10.1007/s10815-012-9813-z. Epub 2012 Jun 13.
- Khosronezhad N, Hassanzadeh V, Hezavehei M, Shahverdi AH, Shahhoseini M. Comparative Epigenetic Analysis of Imprinting Genes Involved in Fertility, in Cryopreserved Human Sperms with Rapid Freezing versus Vitrification Methods. Cell J. 2023 Apr 1;25(4):238-246. doi: 10.22074/cellj.2023.1974291.1171.
- Jenkins TG, Aston KI, Cairns B, Smith A, Carrell DT. Paternal germ line aging: DNA methylation age prediction from human sperm. BMC Genomics. 2018 Oct 22;19(1):763. doi: 10.1186/s12864-018-5153-4.
- Jenkins TG, Aston KI, Carrell DT. Sperm epigenetics and aging. Transl Androl Urol. 2018 Jul;7(Suppl 3):S328-S335. doi: 10.21037/tau.2018.06.10.
- Voros C, Chatzinikolaou F, Papadimas G, Polykalas S, Mavrogianni D, Koulakmanidis AM, Athanasiou D, Kanaka V, Kanaka M, Bananis K, Athanasiou A, Athanasiou A, Papapanagiotou I, Vaitsis D, Tsimpoukelis C, Daskalaki MA, Theodora M, Thomakos N, Antsaklis P, Loutradis D, Daskalakis G. Sperm-Derived Extracellular Vesicles (Sperm-EVs), Emerging Biomarkers and Functional Modulators in Male Infertility and Assisted Reproduction. Genes (Basel). 2025 Nov 22;16(12):1400. doi: 10.3390/genes16121400.
- Fernandez de la Puente M, Salas-Huetos A, Valle-Hita C, Babio N, Murphy MM, Canudas S, Salas-Salvado J. Is telomere length a biomarker of sperm quality? A systematic review and meta-analysis of observational studies. Andrology. 2024 Feb;12(2):277-288. doi: 10.1111/andr.13482. Epub 2023 Jun 25.
- Ferlin A, Rampazzo E, Rocca MS, Keppel S, Frigo AC, De Rossi A, Foresta C. In young men sperm telomere length is related to sperm number and parental age. Hum Reprod. 2013 Dec;28(12):3370-6. doi: 10.1093/humrep/det392. Epub 2013 Oct 27.
- Estudillo E, Jimenez A, Bustamante-Nieves PE, Palacios-Reyes C, Velasco I, Lopez-Ornelas A. Cryopreservation of Gametes and Embryos and Their Molecular Changes. Int J Mol Sci. 2021 Oct 8;22(19):10864. doi: 10.3390/ijms221910864.
- Eisenberg DTA, Lee NR, Rej PH, Hayes MG, Kuzawa CW. Older paternal ages and grandpaternal ages at conception predict longer telomeres in human descendants. Proc Biol Sci. 2019 May 29;286(1903):20190800. doi: 10.1098/rspb.2019.0800. Epub 2019 May 29.
- De Meyer T, Rietzschel ER, De Buyzere ML, De Bacquer D, Van Criekinge W, De Backer GG, Gillebert TC, Van Oostveldt P, Bekaert S; Asklepios investigators. Paternal age at birth is an important determinant of offspring telomere length. Hum Mol Genet. 2007 Dec 15;16(24):3097-102. doi: 10.1093/hmg/ddm271. Epub 2007 Sep 19.
- Chen H, Alves MBR, Belleannee C. Contribution of epididymal epithelial cell functions to sperm epigenetic changes and the health of progeny. Hum Reprod Update. 2021 Dec 21;28(1):51-66. doi: 10.1093/humupd/dmab029.
- Caroppo E, Skinner MK. Could the sperm epigenome become a diagnostic tool for evaluation of the infertile man? Hum Reprod. 2024 Mar 1;39(3):478-485. doi: 10.1093/humrep/dead266.
- Aston KI, Hunt SC, Susser E, Kimura M, Factor-Litvak P, Carrell D, Aviv A. Divergence of sperm and leukocyte age-dependent telomere dynamics: implications for male-driven evolution of telomere length in humans. Mol Hum Reprod. 2012 Nov;18(11):517-22. doi: 10.1093/molehr/gas028. Epub 2012 Jul 9.
Fechas de registro del estudio
Estas fechas rastrean el progreso del registro del estudio y los envíos de resultados resumidos a ClinicalTrials.gov. Los registros del estudio y los resultados informados son revisados por la Biblioteca Nacional de Medicina (NLM) para asegurarse de que cumplan con los estándares de control de calidad específicos antes de publicarlos en el sitio web público.
Fechas importantes del estudio
Inicio del estudio (Actual)
10 de junio de 2026
Finalización primaria (Estimado)
31 de marzo de 2027
Finalización del estudio (Estimado)
30 de septiembre de 2027
Fechas de registro del estudio
Enviado por primera vez
29 de junio de 2026
Primero enviado que cumplió con los criterios de control de calidad
8 de septiembre de 2026
Publicado por primera vez (Actual)
10 de septiembre de 2026
Actualizaciones de registros de estudio
Última actualización publicada (Actual)
10 de septiembre de 2026
Última actualización enviada que cumplió con los criterios de control de calidad
8 de septiembre de 2026
Última verificación
1 de septiembre de 2026
Más información
Términos relacionados con este estudio
Palabras clave
Términos MeSH relevantes adicionales
Otros números de identificación del estudio
- 2303-MUR-046-JM
- 101120126 (Otro número de subvención/financiamiento: European Union Horizon Europe Programme - Marie Skłodowska-Curie Actions Doctoral Network AFRODITA)
Plan de datos de participantes individuales (IPD)
¿Planea compartir datos de participantes individuales (IPD)?
INDECISO
Información sobre medicamentos y dispositivos, documentos del estudio
Estudia un producto farmacéutico regulado por la FDA de EE. UU.
No
Estudia un producto de dispositivo regulado por la FDA de EE. UU.
No
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