Sperm Quality Declines After 2 and 4 Hour Incubation at Room Temperature and 37°C

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Sperm quality declines after 2 and 4 hour incubation at room temperature and 37°C

The Effects of In Vitro Incubation of Asthenoteratozoospermic Semen after Density Gradient Centrifugation at Room Temperature and 37°C on Sperm Parameters, Chromatin Quality and DNA Fragmentation in a Short Time Period

A prospective cohort study, of asthenoteratozoospermia (abnormal sperm motility and morphology) patients, undergoing infertility treatment, was conducted to evaluate the effect of in vitro incubation on prepared sperm samples, at room temperature and 37°C.

A total of twenty seven asthenoteratozoospermic samples were collected and prepared by density gradient centrifugation. The prepared sperm was then separated into 2 parts and incubated at different temperatures. The samples were evaluated before preparation, immediately afterwards and post incubation at 2 and 4 hours for motility, viability, morphology, sperm protamine deficiency, chromatin and DNA fragmentation.

Initial analysis showed a significant increase in both processive motility, total motility and viability following density gradient sperm preparation. This was followed by a significant decrease in processive motility and viability after 2 hours incubation, and again after 4 hours incubation, for both room temperature and 37°C samples.

Interestingly progressive motility and viability of the sperm did not differ significantly between room temperature and 37°C incubation. Significantly morphological analysis found no change in the proportion of normal sperm within the density gradient sperm samples after 0, 2 and 4 hours, at either room temperature or 37°C.

As predicted by similar studies, DNA fragmentation increased at 2 and 4 hours post preparation, however no significant difference was found between the two incubation temperatures, suggesting time post preparation and not temperature, is most important.


  1. Small study size

Similar studies

Ahmed I, et al. (2018). Influence of extended incubation time on Human sperm chromatin condensation, sperm DNA strand breaks and their effect on fertilisation rate. https://doi.org/10.1111/and.12960

Çok T, et al. (2015). The effect of preserving prepared sperm samples at room temperature or at 37°C before intrauterine insemination (IUI) on clinical pregnancy rate. https://dx.doi.org/10.4274/tjod.31644

Thijssen A, et al. (2014). Influence of temperature and sperm preparation on the quality of spermatozoa. Reprod Biomed Online. https://doi.org/10.1016/j.rbmo.2013.12.005

Schuffner A, et al. (2002). Effect of different incubation conditions on phosphatidylserine externalization and motion parameters of purified fractions of highly motile human spermatozoa. https://doi.org/10.1002/j.1939-4640.2002.tb02615.x


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