AgingNCT07814235
Human Sperm Cryopreservation as an Alternative to the Decline of Sperm Quality With Aging
An observational study of Aging and Epigenetic Aging, run by IVI Murcia at 1 site in Spain.
Trial summary
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. Read more about this study
- Who is it for?
- Ages 18 to 65 years, men onlyHealthy volunteers can take part
- Conditions the study is for
- Aging, Epigenetic Aging, Infertility
- How many people
- 45 participants
- Where
- 1 siteSpain
- Study dates
- 10 Jun 2026to 30 Sep 2027
- Phase
- Observational
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 to 65 years
- Men only
- Healthy volunteers accepted
- Adult, Older adult
Study population: 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.
Who may be able to join
- 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
Who cannot join
- 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
Trial locations and contacts
1 site in 1 country. Contact the study team or a site near you to ask about taking part.
Spain1 site, 1 recruiting
- Recruiting
IVI Murcia
Murcia, Murcia, 30007
Juan Carlos Martínez, +34 651 79 10 00, JuanCarlos.Martinez@ivirma.com
About this study
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.
Read the detailed description
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.
- Keywords
- infertility, cryopreservation, sperm, semen, semen analysis, DNA methylation, epigenetics, assisted reproductive technology
- Medical subject headings
- Infertility
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
| Study group | Role | What they receive |
|---|---|---|
| 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. | |
| 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. | |
| 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. |
What is being measured
The primary outcome is the main question the study is designed to answer.
Sperm DNA methylation profile
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.
Measured: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
Secondary outcomes (7)
Sperm concentration
Sperm concentration (million sperm/mL) measured using computer-assisted sperm analysis (CASA).
Measured: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
Total sperm motility
Percentage of motile sperm measured using CASA.
Measured: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
Progressive sperm motility
Percentage of progressively motile sperm measured using computer-assisted sperm analysis (CASA).
Measured: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
Normal sperm morphology
Percentage of sperm with normal morphology assessed by microscopic evaluation of stained slides according to World Health Organization (WHO) criteria.
Measured: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
Sperm DNA fragmentation
DNA fragmentation measured in fresh, slow-frozen, and vitrified sperm samples.
Measured: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
Sperm telomere length
Sperm telomere length measured in fresh, slow-frozen, and vitrified sperm samples.
Measured: Baseline (study semen collection visit) and after thawing of cryopreserved semen samples, through study completion (up to 9 months).
Seminal plasma extracellular vesicle composition
Characterization of seminal plasma extracellular vesicle composition in fresh semen samples.
Measured: Baseline (study semen collection visit).
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted10 Sep 2026
- Study start10 Jun 2026
- Primary completion31 Mar 2027 (estimated)
- Study completion30 Sep 2027 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (28)Papers the sponsor has linked to this study.
- Other trials for AgingStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Cohort
- Time perspective
- Prospective
- Enrolment
- 45 participants (estimated)
Sponsor and investigators
- Lead sponsor
- IVI Murcia
- Sponsor type
- University, hospital or other
- Collaborators
- European Union
- Responsible party
- Sponsor
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT07814235
- Sponsor's protocol ID
- 2303-MUR-046-JM
- Other IDs
- 101120126
- Status last verified
- Sep 2026
- First submitted
- 29 Jun 2026
- Last updated
- 10 Sep 2026
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- Undecided
Publications
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.
PubMed 35805194Wang 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.
PubMed 34712227Tao 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.
PubMed 32145746Sysoeva 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.
PubMed 34855079Khosravizadeh 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.
PubMed 35713751Sciamanna 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.
PubMed 31455195Reshi 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.
PubMed 36469292Practice 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.
PubMed 23391408Njajou 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.
PubMed 17623782Machtinger 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.
PubMed 26663221Lismer 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.
PubMed 37059740Laurentino 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.
PubMed 32951333Kumar 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.
PubMed 34277615Klaver 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.
PubMed 22692281Khosronezhad 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.
PubMed 37210644