Human Spermatozoa Ultrastructure Assessment in the Infertility Treatment by Assisted Reproduction Technique

Abstract

The aim of this study was to prospectively investigate the spermatozoa ultrastructure in relation to the results of in vitro fertilization -embryo transer (IVF-ET).Forty-nine consecutive couples admitted for IVF-ET were prospectively evaluated for electron microscopic spermatozoa morphology and the outcome of IVF-ET. Thirty-four couples revealed successful fertilization, defined as presence of two pronuclei 14-16 hours after spermatozoa administration, while the remaining 15 formed the failure group. Spermatozoa fixed with 2.5% glutaraldehyd and embedded in Spurr's resin were analyzed with JAM 100 S transmission electron microscope (TEM) for the following ultrastructure abnormalities: head deformity, cytoplasmic residues, chromatin condensation failures, acrosomal alterations, neck defects, midpiece defects, principal piece and end-piece defects and immature forms.Successful IVF-ET couples revealed a significantly higher percentage of normal spermatozoa utrastructure (32.0 AE 13.1% versus 17.1 AE 13.4%, p < 0.001). Failed IVF-ET couples represented a significantly higher percentage of chromatin condensation failures (9.8 AE 5.1% versus 5.7 AE 5.3%, p < 0.05) and tail defects (16.7 AE 11.5% versus 7.2 AE 7.2%, p < 0.001). A positive correlation between normal ultrastructure spermatozoa percentage and fertilized oocytes percentage was found (r ¼ 0.35, p < 0.05).Our data suggest that spermatozoa TEM findings correlate with IVF-ET results. Ultrastructural estimation of spermatozoa can improve the diagnosis of male fertility and may explain some reasons of failure in assisted reproduction methods. We consider systematic TEM spermatozoa examination in cases with failed IVF-ET prior to intracytoplasmic sperm injection (ICSI). KEYWORDS in vitro fertilization, spermatozoa, ultrastructureAbbreviations: IVF-ET: in vitro fertilization -embryo transfer; TEM: transmission electron microscope; ICSI intracytoplasmic sperm injection.

Combination of density gradient centrifugation and swim-up methods effectively decreases morphologically abnormal sperms

“…Ng et al [19] revealed that sperms with good motility were effectively enriched when the methods were combined compared to the swim-up method alone; however, they did not report the elimination of sperms with ultrastructural abnormal morphologies. Ultrastructural analysis of morphology using a transmission electron microscope (TEM) can detect sperm abnormalities more precisely than a light microscope (LM), and IVF outcomes can be predicted based on the rate of ultrastructural sperm abnormalities [20, 21]. Therefore, analysis of the fine structure of sperms using TEM may lead to the establishment of a sperm washing protocol for sorting good-quality sperms.…”

Effect of systemic isotretinoin on semen parameters and sperm ultrastructure in male patients treated for acne vulgaris : An electron microscope study

“…TEM is the most accurate method assessing teratozoospermia in males either fertile or not; heterogeneous alterations in the shape of different sperm components that affect the fertilizing capacity of male patient can be discovered by electron microscope despite normal appearance by light microscope, which may reflect the functional and structural disorders of the altered sperms [29] . Ultrastructural defects of spermatozoa by electron microscope had been studied by many authors, and different classifications were taken into consideration [17,[30][31][32] .…”

“…In this study, we had used computer-assisted semen analysis, and in addition, we were the first to use TEM as a precise method to study sperm ultrastructure, sperm structure dimensions , and sperm morphology before and after treatment to study accurately if systemic isotretinoin had a harmful effect on male fertility potential or not. Kotwicka et al [17] studied sperm ultrastructure changes in the 49 infertile couples undergoing in vitro fertilization technique. They reported eight ultrastructural parameters to be measured by TEM in spermatozoa of infertile men, which included acrosomal alterations, chromatin condensation failure, cytoplasmic residues, head deformity, neck defects, midpiece defects, main piece and end-piece tail defects, and immature forms.…”

“…To assess sperm ultrastructure changes by TEM, at least 80 complete undisrupted sperms were examined for structural defects. Findings were counted and divided into the following categories: normal forms, head deformity, cytoplasmic residues, chromatin condensation failures, acrosomal alterations, neck defects, midpiece defects, principal piece and end-piece defects, and immature forms [17] . The mean frequencies of these parameters were measured for every patient before and after isotretinoin treatment (Fig.…”

Protective Role of Purified Cysteine Proteinases against Fasciola gigantica Infection in Experimental Animals

“…However, despite long-standing research, a vaccine against this parasite has not yet been developed to the point of commercialization [17]. This can be largely attributed to a fact that immune responses to vaccines are influenced by the route of immunization (injection or oral), form of antigen, and presence of adjuvant in the vaccine [18,19].…”