Abstract
The acrosome of the spermatozoa is required for fertilization and in the raw ejaculate the percentage of viable acrosome-intact spermatozoa, the acrosomal status, is higher among men with good semen quality. Here we investigated if the acrosomal status of the processed semen preparations used at a fertility clinic can also be informative and whether it is associated with fecundity. The acrosomal status was measured by image cytometry on purified semen samples from couples during in vitro fertilization (IVF) (n = 99) and intracytoplasmic sperm injection (ICSI) (n = 107) treatment. Purified frozen-thawed donor samples were also analyzed (n = 199). In purified semen preparations the acrosomal status was significantly higher among sperm donors (p = 5.3 × 10−8) and men from IVF couples (p = 2.2 × 10−5) when compared to men from ICSI couples. A significant difference was also found between female, male and mixed factor infertility (p = 0.003). No association with lifestyle factors was found. In frozen-thawed donor samples, a significant positive (r = 0.16, p = 0.025) association with the number of pregnancies per sold straw was observed together with an area under the curve of 75.3%, when comparing the top and bottom deciles. Our results indicate that the acrosomal status may be a valuable parameter for personalizing fertility treatments and might be a good predictor of pregnancy success among normozoospermic men.
Highlights
The number of couples seeking help for infertility is increasing worldwide [1,2,3]
Evaluation of semen quality is important in the clinical work-up of infertile couples and contributes to the clinical decision tree for choosing the type of treatment, i.e., intrauterine insemination (IUI), in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI)
The spermatozoa pellet was resuspended in a staining solution containing: 5 μg/mL fluorescein isothiocyanate conjugated Pisum sativum agglutinin (FITC-Pisum Sativum Agglutinin (PSA), Sigma-Aldrich, Soeborg, Denmark), 0.5 μg/mL propidium iodide (PI, ChemoMetec A/S, Alleroed, Denmark), and 10 μg/mL Hoechst-33342 (H342, ChemoMetec, Alleroed, Denmark) in PBS and incubated for 30 min at 37 ◦C
Summary
The number of couples seeking help for infertility is increasing worldwide [1,2,3]. In 2018, approximately 1 out of 10 Danish children was born after fertility treatment [4]. We have developed an assay based on image cytometry, which allows automatization and easy determination of the percentage of viable acrosome-intact spermatozoa in an ejaculate, an indicator of the acrosomal status [15,16] This assay exploits the advantage of PSA coupled to fluorescein isothiocyanate (FITC) to flow via membrane pores into the acrosomal compartment of reacting acrosomes and to stabilize the acrosomal matrix allowing easy detection of spermatozoa undergoing the acrosome reaction [17]. The objective of this study was to investigate the acrosomal status of the spermatozoa directly used in fertility treatment, i.e., density gradient purified semen preparations from infertile couples and purified-frozen-thawed preparations from sperm donors. We further aimed to test whether the acrosomal status could be associated with lifestyle factors of the infertile couples and fecundity among semen donors
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