Semen preparation methods and sperm apoptosis: swim-up versus gradient-density centrifugation technique

Ricci, Giuseppe; Perticarari, Sandra; Boscolo, Rita; Montico, Marcella; Guaschino, Secondo; Presani, Gianni

doi:10.1016/j.fertnstert.2007.11.068

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Volume 91, Issue 2 , Pages 632-638, February 2009

Semen preparation methods and sperm apoptosis: swim-up versus gradient-density centrifugation technique

Giuseppe Ricci, M.D.
- Assisted Reproduction Unit, Department of Obstetrics and Gynecology, Institute of Child Health IRCCS Burlo Garofolo, and University of Trieste, Trieste, Italy
- Reprint requests: Giuseppe Ricci, M.D., Assisted Reproduction Unit, Department of Obstetrics and Gynecology, Institute of Child Health IRCCS Burlo Garofolo and University of Trieste, Via dell'Istria, 65/1, 34137, Trieste, Italy (FAX: +39 (0) 40 76 12 66).
Sandra Perticarari, B.Sc.
- Clinical Analysis Unit, Department of Laboratory Medicine, Institute of Child Health IRCCS Burlo Garofolo, and University of Trieste, Trieste, Italy
Rita Boscolo, B.Sc.
- Assisted Reproduction Unit, Department of Obstetrics and Gynecology, Institute of Child Health IRCCS Burlo Garofolo, and University of Trieste, Trieste, Italy
Marcella Montico, B.Sc.
- Epidemiology Unit, Institute of Child Health IRCCS Burlo Garofolo, and University of Trieste, Trieste, Italy
Secondo Guaschino, M.D.
- Assisted Reproduction Unit, Department of Obstetrics and Gynecology, Institute of Child Health IRCCS Burlo Garofolo, and University of Trieste, Trieste, Italy
Gianni Presani, B.Sc.
- Clinical Analysis Unit, Department of Laboratory Medicine, Institute of Child Health IRCCS Burlo Garofolo, and University of Trieste, Trieste, Italy

Received 2 September 2007; received in revised form 19 November 2007; accepted 19 November 2007. published online 18 January 2008.

Objective

To compare the effects of density-gradient centrifugation and swim-up on sperm apoptosis by using a multiparameter flow cytometric method.

Design

Autocontrolled split-sample study.

Setting

Tertiary infertility center.

Patient(s)

Sixty-two male partners of couples undergoing infertility investigations.

Intervention(s)

Each sample was analyzed both before and after semen preparation by optical microscopy and by flow cytometry.

Main Outcome Measure(s)

Percentage of viable, apoptotic, and necrotic sperm and recovery rate of total motile, progressive motile, and viable sperm before and after the two sperm preparation methods.

Result(s)

Compared with the original semen, the mean percentages of apoptotic and necrotic sperm were significantly lower after both sperm preparation methods. The mean percentage of viable sperm was significantly higher after swim-up compared with gradient centrifugation. The recovery rates of total motile, progressive motile, and viable sperm were significantly higher using gradient centrifugation compared with swim-up. The viable sperm percentage and the progressive sperm motility were significant predictors for negative difference between the two methods in terms of viable sperm percentage after preparation.

Conclusion(s)

Both sperm preparation methods allow obtaining a sperm population with a low percentage of apoptotic sperm. Therefore, the risk of using apoptotic sperm for clinical treatment seems to be rather low. The choice of method will depend on whether IVF/ICSI or intrauterine insemination is to be performed.

Key Words: Apoptosis, density-gradient centrifugation, flow cytometry, fluorospheres, semen analysis, sperm, swim-up, Syto 16