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Spermatogenesis is the process by which haploid sperm cells are produced in the male testes. It starts with stem cells located close to the outer rim of seminiferous tubules. These spermatogonial stem cells divide asymmetrically to give rise to additional stem cells (meaning that these structures “self-renew”), as well as sperm progenitors, called spermatocytes. Importantly, this method of asymmetric mitotic division maintains a population of spermatogonial stem cells in the male...
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During ejaculation, males release around 2-5 milliliters of semen, which is a complex mixture of mature sperm and various fluids produced by accessory glands. The mature sperm cells measure approximately 60 micrometers in length and consist of a head, neck, midpiece, and tail. The head is flattened and tapered, measuring about 4 to 5 micrometers in length. It contains a nucleus with condensed chromosomes and an acrosome, a cap-like structure filled with enzymes essential for penetrating the...
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Related Experiment Video

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Sperm Collection of Differential Quality Using Density Gradient Centrifugation
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The Ticking Clock: Differential Time-Dependent Deterioration Between Washed and Thawed Sperm.

Adiel Kahana1, Emily Hamilton2, Noga Fuchs Weizman1,3

  • 1The Institute for the Study of Fertility and Racine IVF Unit, Lis Maternity Hospital, Tel Aviv Sourasky Medical Center, Tel Aviv, Israel.

Andrology
|November 25, 2025
PubMed
Summary

Thawed sperm deteriorates faster than fresh sperm in key reproductive parameters. Immediate use after thawing is crucial for optimizing outcomes in assisted reproduction.

Keywords:
DNA fragmentationintrauterine inseminationsperm cryopreservationsperm washingtime‐dependent deterioration

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Area of Science:

  • Reproductive Biology
  • Cryobiology
  • Spermatozoa Research

Background:

  • Cryopreservation is standard in assisted reproductive technologies.
  • The comparative time-dependent deterioration rate between fresh and thawed sperm remains unclear.

Purpose of the Study:

  • To directly compare the deterioration rates of sperm motility, vitality, and DNA fragmentation between fresh washed and thawed sperm over time.
  • To assess the impact of cryopreservation and thawing processes on sperm quality.

Main Methods:

  • Prospective study involving 50 males, splitting samples into fresh washed and thawed groups.
  • Sperm parameters (motility, vitality, DNA fragmentation index) assessed at baseline and after 75 min incubation.
  • Control experiments evaluated cryoprotectant effects and post-thaw washing impact.

Main Results:

  • Thawed sperm showed significantly greater declines in motility (29% vs 17%) and vitality (21% vs 7%) compared to fresh samples (p < 0.0001).
  • DNA fragmentation index increased significantly in thawed samples, reaching critical levels (mean 34%), unlike fresh samples (mean 12%).
  • Cryoprotectant exposure alone did not cause deterioration, and post-thaw washing offered no improvement.

Conclusions:

  • Thawed sperm exhibits accelerated deterioration across motility, vitality, and DNA integrity due to cryopreservation stress.
  • Minimizing the time between thawing and insemination is essential for improving reproductive success.
  • Immediate use of thawed spermatozoa post-thaw is recommended to optimize assisted reproductive outcomes.