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Comparison of two-layer Percoll gradient and mini-Percoll methods for sperm preparation
S Kunathikom1, S Ko-Anantakul, K Techatraisak
1Department of Obstetrics and Gynecology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Abstract:
To compare the efficiency of sperm preparation between the two-layer Percoll gradient and mini-Percoll methods, 50 normal and 33 abnormal semen samples from male partners of infertile couples were studied. The number of recovered spermatozoa, percentage of motility, percentage of normal morphology, and their survival at 24 and 48 hours were assessed. Both Percoll gradient techniques resulted in a significantly higher percentage of motility and percentage of normal morphology compared with the original semen samples (p < 0.0001). The two-layer Percoll gradient showed a higher sperm recovery than the mini-Percoll method (p < 0.001), but the latter resulted in a higher percentage of motility (p > 0.001) and a higher sperm survival rate at 24 hours (p < 0.05) than the former, regarding normal semen samples. These differences did not appear with abnormal semen samples when analyzed as a group. Considering each of the abnormal parameters separately, sperm recovery was significantly higher after the two-layer Percoll gradient in the case of astheno- and teratozoospermia (p < 0.05), but sperm survival at 48 hours was higher after the mini-Percoll gradient in the case of teratozoospermia (p < 0.05). It is concluded that both the two-layer Percoll gradient and mini-Percoll method can be used effectively for sperm preparation. The former yields a higher sperm recovery, but the latter should be considered regarding teratozoospermic samples and semen samples of very low volume.
Insights
Both two-layer Percoll gradient and mini-Percoll methods effectively prepare sperm. Mini-Percoll offers better motility and survival for normal samples, while two-layer Percoll provides higher recovery for specific abnormal semen parameters.
Area of Science:
- Reproductive Medicine
- Andrology
- Sperm Preparation Techniques
Background:
- Sperm preparation is crucial for assisted reproductive technologies.
- Percoll gradients are widely used for sperm separation and purification.
- Comparing different Percoll-based methods is essential for optimizing outcomes.
Purpose of the Study:
- To compare the efficiency of two-layer Percoll gradient and mini-Percoll methods for sperm preparation.
- To evaluate sperm recovery, motility, morphology, and survival rates using both techniques.
Main Methods:
- Analysis of 50 normal and 33 abnormal semen samples.
- Comparison of sperm parameters (recovery, motility, morphology, survival) after preparation with two-layer Percoll and mini-Percoll.
- Statistical analysis to determine significant differences between methods and sample types.
Main Results:
- Both methods significantly improved sperm motility and normal morphology compared to original samples.
- Two-layer Percoll yielded higher sperm recovery (p < 0.001).
- Mini-Percoll showed higher motility (p > 0.001) and 24-hour survival (p < 0.05) in normal samples; specific advantages noted for teratozoospermia and asthenozoospermia.
Conclusions:
- Both two-layer Percoll gradient and mini-Percoll are effective for sperm preparation.
- Two-layer Percoll offers superior sperm recovery.
- Mini-Percoll is advantageous for teratozoospermic samples and low-volume semen, particularly regarding motility and short-term survival.