Comparison of two sperm processing techniques for low complexity assisted fertilization: sperm washing followed by swim-up and discontinuous density gradient centrifugation.

Cássio L Fácio,Ligiane A Machado-Paula,Edilberto Araújo Filho,Lígia F Previato,Paulo CS Matheus

doi:10.5935/1518-0557.20160040

Cássio L Fácio, Ligiane A Machado-Paula + Show 3 more

Open Access

https://doi.org/10.5935/1518-0557.20160040

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Journal: JBRA assisted reproduction	Publication Date: Jan 1, 2016
Citations: 26	License type: CC BY 4.0

Abstract

ObjectiveThis study aimed to assess and compare sperm motility, concentration, and morphology recovery rates, before and after processing through sperm washing followed by swim-up or discontinuous density gradient centrifugation in normospermic individuals.MethodsFifty-eight semen samples were used in double intrauterine insemination procedures; 17 samples (group 1) were prepared with sperm washing followed by swim-up, and 41 (group 2) by discontinuous density gradient centrifugation. This prospective non-randomized study assessed seminal parameters before and after semen processing. A dependent t-test was used for the same technique to analyze seminal parameters before and after semen processing; an independent t-test was used to compare the results before and after processing for both techniques.ResultsThe two techniques produced decreases in sample concentration (sperm washing followed by swim-up: P<0.000006; discontinuous density gradient centrifugation: P=0.008457) and increases in motility and normal morphology sperm rates after processing. The difference in sperm motility between the two techniques was not statistically significant. Sperm washing followed by swim-up had better morphology recovery rates than discontinuous density gradient centrifugation (P=0.0095); and the density gradient group had better concentration recovery rates than the swim-up group (P=0.0027).ConclusionThe two methods successfully recovered the minimum sperm values needed to perform intrauterine insemination. Sperm washing followed by swim-up is indicated for semen with high sperm concentration and better morphology recovery rates. Discontinuous density gradient centrifugation produced improved concentration recovery rates.

Highlights

Spermatozoa undergo a series of biochemical and structural changes within the female genital tract called capacitation (Neves, 1991)
The 58 semen samples were divided into two groups: group 1 with 17 samples submitted to sperm washing followed by swim-up; and group 2, with 41 samples submitted to discontinuous density gradient centrifugation
The assessment of each technique in terms of semen parameters before and after processing revealed that both resulted in decreased sperm concentration after processing (P=0.000006 for sperm washing followed by swim-up; and P=0.008457 for discontinuous density gradient centrifugation), and increased motility and rates of morphologically normal sperm (P=0.00001 for sperm washing followed by swim-up; and P

Summary

Introduction

Spermatozoa undergo a series of biochemical and structural changes within the female genital tract called capacitation (Neves, 1991). In-vivo, capacitation occurs over a period of seven hours (Yoshida et al, 2008). During this period, the glycoprotein coat and seminal proteins are removed from the surface of the sperm’s acrosome. Capacitated spermatozoa show highly active flagellar beating (hyperactivation), undergo the acrosome reaction, penetrate the pellucid zone, and bind and fuse with the oocytes (Yoshida et al, 2008). Sperm capacitation in assisted human reproduction (AHR) is performed artificially using specific techniques. Intrauterine insemination (IUI), an assisted reproduction technology (ART), is a non-invasive method and the least expensive of ARTs (Dodson & Haney, 1991). Seminal parameters significantly impact the outcome of IUI (Arny & Quagliarello, 1987; Brasch et al 1994; Berg et al, 1997; Dorjpurev et al, 2011)

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