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Studies on the basic issues relevant to sperm cryopreservation in humans.

Huanhuan Hu1, Xiaowei Shi1, Guojie Ji1

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|June 11, 2020
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Summary

Optimizing human sperm cryopreservation involves using specific freezing media osmolality and sugars like trehalose. Rapid freezing with trehalose at 37°C thawing yields the best sperm motility and DNA integrity.

Keywords:
humanrapid freezingspermsucrosetrehalose

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Area of Science:

  • Reproductive Biology
  • Cryobiology
  • Andrology

Background:

  • Human sperm cryopreservation is crucial for assisted reproduction technologies.
  • Optimizing rapid freezing protocols is essential for maintaining sperm viability and function.
  • Key factors influencing cryopreservation success, including medium osmolality, sperm concentration, thawing methods, and cryoprotective sugars, require further investigation.

Purpose of the Study:

  • To investigate the impact of freezing medium osmolality, sperm concentration, thawing temperature, and the type of sugar (sucrose vs. trehalose) on human sperm motility and DNA integrity.
  • To determine optimal parameters for rapid freezing of human sperm using standard 0.5 ml straws.

Main Methods:

  • Human sperm samples were cryopreserved using rapid freezing in 0.5 ml straws.
  • Variables tested included freezing medium osmolality (442 vs. 536 mOsm/kg), sperm concentrations (5-20 × 10^6 sperm/ml), thawing methods (37°C for 2 min vs. 42°C for 15 s), and cryoprotective sugars (0.25 M sucrose vs. 0.25 M trehalose).
  • Post-thaw sperm motility and DNA integrity were assessed.

Main Results:

  • Lower osmolality (442 mOsm/kg) freezing medium resulted in significantly higher post-thaw total and progressive motility compared to higher osmolality (536 mOsm/kg).
  • Trehalose (0.25 M) demonstrated superior cryoprotective effects on sperm motility compared to sucrose (0.25 M).
  • Thawing at 37°C for 2 minutes was significantly more effective than thawing at 42°C for 15 seconds for preserving sperm motility. Sperm DNA integrity was relatively unaffected by these factors.

Conclusions:

  • Optimal human sperm cryopreservation via rapid freezing is achieved using 0.25 M trehalose or sucrose in a freezing medium with an osmolality of 442-457 mOsm/kg, high sperm concentration, and thawing at 37°C.
  • Trehalose serves as a more potent cryoprotectant for human sperm than sucrose.
  • Sperm motility is more sensitive to cryopreservation conditions than sperm DNA integrity.