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Purpose <p>To evaluate whether long-term cryostorage duration affects post-thaw sperm motility and progressive motile concentration (PMC) in samples cryopreserved for up to 30 years.</p> Methods <p>A retrospective cohort study of 629 donor sperm samples cryopreserved between 1990 and 2020 in a single university-affiliated andrology laboratory was conducted. Motility and PMC were assessed immediately after thawing (T0) and after extended storage (TES). Associations between storage duration and post-thaw changes were evaluated using correlation analysis, multivariable linear regression, and multilevel mixed-effects models with donor-level random effects. K-means clustering was applied to identify sperm quality phenotypes.</p> Results <p>Total motility declined significantly following the freeze–thaw process (<i>p</i> &lt; 0.001) but remained within clinically usable ranges after extended storage. Storage duration showed only a weak association with relative motility change (<i>r</i> = 0.169, <i>p</i> &lt; 0.001), without a consistent duration-dependent trend in absolute post-thaw motility. PMC changes were modest and non-monotonic across storage groups. Multilevel models indicated that storage duration explained only a small proportion of variance in long-term outcomes. Cluster analysis identified distinct sperm quality phenotypes independent of storage duration.</p> Conclusions <p>Long-term cryostorage of up to 30 years is not associated with substantial impairment of sperm motility or PMC. These findings support the stability of cryopreserved spermatozoa under prolonged storage conditions.</p>

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Long-term cryostorage does not impair human sperm motility: evidence from a three-decade cohort study

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月7日

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  • Shimi Barda,
  • Shanee Dim,
  • Shlomit Shabat,
  • Sandra Edith Kleiman,
  • Noga Fuchs Weizman

摘要

Purpose

To evaluate whether long-term cryostorage duration affects post-thaw sperm motility and progressive motile concentration (PMC) in samples cryopreserved for up to 30 years.

Methods

A retrospective cohort study of 629 donor sperm samples cryopreserved between 1990 and 2020 in a single university-affiliated andrology laboratory was conducted. Motility and PMC were assessed immediately after thawing (T0) and after extended storage (TES). Associations between storage duration and post-thaw changes were evaluated using correlation analysis, multivariable linear regression, and multilevel mixed-effects models with donor-level random effects. K-means clustering was applied to identify sperm quality phenotypes.

Results

Total motility declined significantly following the freeze–thaw process (p < 0.001) but remained within clinically usable ranges after extended storage. Storage duration showed only a weak association with relative motility change (r = 0.169, p < 0.001), without a consistent duration-dependent trend in absolute post-thaw motility. PMC changes were modest and non-monotonic across storage groups. Multilevel models indicated that storage duration explained only a small proportion of variance in long-term outcomes. Cluster analysis identified distinct sperm quality phenotypes independent of storage duration.

Conclusions

Long-term cryostorage of up to 30 years is not associated with substantial impairment of sperm motility or PMC. These findings support the stability of cryopreserved spermatozoa under prolonged storage conditions.