Human Sperm Cryopreservation as an Alternative to the Decline of Sperm Quality With Aging
8. september 2026 oppdatert av: 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.
Studieoversikt
Status
Rekruttering
Detaljert beskrivelse
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.
Studietype
Observasjonsmessig
Registrering (Antatt)
45
Kontakter og plasseringer
Denne delen inneholder kontaktinformasjon for de som utfører studien, og informasjon om hvor denne studien blir utført.
Studiesteder
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Murcia
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Murcia, Murcia, Spania, 30007
- Rekruttering
- IVI Murcia
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Ta kontakt med:
- Juan Carlos Martínez
- Telefonnummer: +34 651 79 10 00
- E-post: JuanCarlos.Martinez@ivirma.com
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Deltakelseskriterier
Forskere ser etter personer som passer til en bestemt beskrivelse, kalt kvalifikasjonskriterier. Noen eksempler på disse kriteriene er en persons generelle helsetilstand eller tidligere behandlinger.
Kvalifikasjonskriterier
Alder som er kvalifisert for studier
- Voksen
- Eldre voksen
Tar imot friske frivillige
Ja
Prøvetakingsmetode
Ikke-sannsynlighetsprøve
Studiepopulasjon
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.
Beskrivelse
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
Studieplan
Denne delen gir detaljer om studieplanen, inkludert hvordan studien er utformet og hva studien måler.
Hvordan er studiet utformet?
Designdetaljer
Kohorter og intervensjoner
Gruppe / Kohort |
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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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Hva måler studien?
Primære resultatmål
Resultatmål |
Tiltaksbeskrivelse |
Tidsramme |
|---|---|---|
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Sperm DNA methylation profile
Tidsramme: 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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Sekundære resultatmål
Resultatmål |
Tiltaksbeskrivelse |
Tidsramme |
|---|---|---|
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Sperm concentration
Tidsramme: 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
Tidsramme: 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
Tidsramme: 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
Tidsramme: 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
Tidsramme: 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
Tidsramme: 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
Tidsramme: 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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Andre resultatmål
Resultatmål |
Tiltaksbeskrivelse |
Tidsramme |
|---|---|---|
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Blood leukocyte telomere length
Tidsramme: 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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Samarbeidspartnere og etterforskere
Det er her du vil finne personer og organisasjoner som er involvert i denne studien.
Sponsor
Samarbeidspartnere
Publikasjoner og nyttige lenker
Den som er ansvarlig for å legge inn informasjon om studien leverer frivillig disse publikasjonene. Disse kan handle om alt relatert til studiet.
Generelle publikasjoner
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- 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.
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- 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.
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- 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.
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- 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.
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Studierekorddatoer
Disse datoene sporer fremdriften for innsending av studieposter og sammendragsresultater til ClinicalTrials.gov. Studieposter og rapporterte resultater gjennomgås av National Library of Medicine (NLM) for å sikre at de oppfyller spesifikke kvalitetskontrollstandarder før de legges ut på det offentlige nettstedet.
Studer hoveddatoer
Studiestart (Faktiske)
10. juni 2026
Primær fullføring (Antatt)
31. mars 2027
Studiet fullført (Antatt)
30. september 2027
Datoer for studieregistrering
Først innsendt
29. juni 2026
Først innsendt som oppfylte QC-kriteriene
8. september 2026
Først lagt ut (Faktiske)
10. september 2026
Oppdateringer av studieposter
Sist oppdatering lagt ut (Faktiske)
10. september 2026
Siste oppdatering sendt inn som oppfylte QC-kriteriene
8. september 2026
Sist bekreftet
1. september 2026
Mer informasjon
Begreper knyttet til denne studien
Nøkkelord
Ytterligere relevante MeSH-vilkår
Andre studie-ID-numre
- 2303-MUR-046-JM
- 101120126 (Annet stipend/finansieringsnummer: European Union Horizon Europe Programme - Marie Skłodowska-Curie Actions Doctoral Network AFRODITA)
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Studerer et amerikansk FDA-regulert medikamentprodukt
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Nei
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