Silane-coated silica particle solutions (ISolate(TM) and PureSperm)TM)) and iodixanol (OptiPrep(TM)) were compared to polyvinylpyrrolidone (PVP)-coated silica particles (Percoll(TM)) in their efficacy to recover spermatozoa by gradient centrifugation for use in assisted reproductive procedures. Efficacy was assessed in terms of percentages of sperm recovery, sperm vitality and motility, normal sperm morphology and normal sperm chromatin condensation. No significant difference was found in the recovery of spermatozoa for men with both normal sperm counts and oligozoospermia, between PVP-coated and silane-coated particle solutions. Iodixanol had significantly lower sperm recovery compared to the other products. Sperm vitality, progressive motility, normal morphology and normal chromatin condensation did not differ significantly between any of the sperm isolation products.
The human sperm motility assay was used as a measure of quality control in the IVF laboratory. The effects of albumin supplementation and incubation time on the sensitivity of the human sperm motility assay were investigated. The assay was also compared with mouse embryo development. The human sperm motility assay and mouse embryo development assays were performed on 25 items commonly used in IVF laboratories. Sperm motility assay was conducted after 2, 4, 6, 8, 24 and 48 h incubation intervals under standard embryology conditions. A calculated sperm motility index value <0.75 was used to indicate sperm toxicity. It was found that optimum sensitivity (P < 0.01) of the human sperm motility assay was attained in the absence of albumin after 4, 8 and 48 h incubation periods. Items identified to be sperm toxic within 8 h by the human sperm motility assay were considered to be of clinical significance due to the close concordance of these results with mouse embryo development.
The prediction of human fertilization is an important aspect of research protocols dealing with male fertility. Sperm DNA has been reported to be an indicator of human sperm fertility potential. The Acridine Orange test for evaluation of sperm DNA has been employed during the present study to determine its relationship with human sperm morphology and fertilization in vitro. Seventy-six patients from the in-vitro fertilization (IVF) and/or gamete intra-Fallopian transfer (GIFT) programme were randomly selected for the study. All patients had a routine semen analysis, sperm DNA evaluation and underwent standard IVF procedures at the time of the study. The results indicated a moderate positive correlation (r = 0.38, P = 0.0006) between results of the Acridine Orange test and normal sperm morphology. Patients with an Acridine Orange test value exceeding 24% had significantly higher oocyte fertilization rates than patients with lower values, for metaphase I (74 versus 51%, P = 0.0008) and for metaphase II oocytes (88 versus 60%, P = 0.0001). Sperm morphology, however, proved to be a more significant predictor of fertilization in vitro compared to the Acridine Orange test.
The aim of this study was twofold: to investigate the ability of Percoll gradient centrifugation (52, 68, 84%) to fractionate semen samples according to motility quality and percentage normal morphology and to determine whether there is an association between sperm motility quality and percentage normal morphology. Sperm motility was evaluated using a Hamilton Thorn analyzer and normal sperm morphology was manually assessed according to the strict criteria (< or = 4, 5-14, and >14%). The majority of motility parameters and the percentage normal morphology were found to be significantly improved in the 84% Percoll fraction. The greatest effect was on the < or = 4% group, shifting the mean normal morphology percentage from 2.6 to 5.2%. Curvilinear velocity (VCL) and average path velocity (VAP) were the only two motility parameters that were significantly associated with the percentage normal morphology. Using a combined VCL, VAP vector the >14% group was found to have a significantly different value as compared to the 5-14 and < or = 4% groups. Percoll (discontinuous) gradient centrifugation can therefore play a significant role in the improvement of semen samples for use in assisted reproduction procedures. The VCL, VAP vector identified may also serve as an additional tool in the prediction of the fertility potential of sperm samples.
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