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Updated: Jun 18, 2026
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Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects
Published on: April 21, 2022
Leukocytospermia and sperm preparation--a flow cytometric study
Giuseppe Ricci1, Sandra Perticarari, Rita Boscolo
1Assisted Reproduction Unit, Department of Obstetrics and Gynaecology, Institute for Maternal and Child Health IRCCS Burlo Garofolo and University of Trieste, Via dell'Istria 65/1, Trieste 34137, Italy. ricci@burlo.trieste.it
Reproductive Biology and Endocrinology : RB&E
|November 21, 2009
Summary
Swim-up method is more effective than density-gradient centrifugation in reducing seminal leukocytes. Leukocyte levels do not impact sperm preparation, but swim-up is preferred for assisted conception. Peroxidase test is unreliable for detecting leukocytes.
Area of Science:
- Reproductive Biology
- Andrology
- Clinical Laboratory Science
Background:
- Leukocytes in semen are a primary source of reactive oxygen species, potentially impairing sperm function and fertilization rates in assisted reproduction.
- The World Health Organization (WHO) recommends the peroxidase test for detecting seminal leukocytes, but its accuracy is questionable.
- Understanding leukocyte impact and optimizing their removal are crucial for improving assisted reproductive technologies (ART).
Purpose of the Study:
- To compare the efficacy of swim-up and density-gradient centrifugation in removing seminal leukocytes.
- To evaluate the influence of leukocytes on sperm preparation outcomes.
- To compare flow cytometry with the peroxidase test for quantifying leukocyte concentration in semen.
Main Methods:
- Analyzed semen samples from 126 men undergoing infertility investigations for leukocytospermia using microscopy and flow cytometry.
- Processed 69 samples using both swim-up and density-gradient centrifugation techniques.
- Utilized multiparameter flow cytometry to assess sperm concentration, viability, apoptosis, and leukocyte concentration in neat and prepared sperm.
Main Results:
- Both sperm preparation methods significantly reduced leukocyte concentrations, with swim-up showing a more substantial decrease than density-gradient centrifugation.
- Initial or prepared leukocyte concentrations did not correlate with post-processing sperm parameters.
- A significant correlation was observed between residual leukocytes in prepared fractions and viable sperm recovery rate; flow cytometry demonstrated higher sensitivity than the peroxidase test.
Conclusions:
- Seminal leukocytes do not appear to negatively affect sperm preparation results.
- The swim-up method is recommended for processing semen samples with leukocytes in assisted conception.
- The peroxidase test is not suitable for clinical research due to its low sensitivity in detecting leukocytospermia.
