• Clinical and Experimental Reproductive Medicine
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• v.41 no.3
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• pp.132-136
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• 2014

Objective: The presence of sperm-head vacuoles has been suspected to be deleterious to the outcomes of assisted reproductive technology (ART). It is difficult to accurately distinguish morphologically abnormal sperm with vacuoles under a light microscope. This study was performed to analyze the result of the observation of sperm-head vacuoles using Papanicolaou staining under a light microscope and whether the male partner's age affects these vacuoles. Methods: Sperm morphology with vacuoles was evaluated using Papanicolaou staining and observed under a light microscope ($400{\times)$) in 980 men. The normal morphology was divided into three categories (group A, <4% of normal morphology; group B, 4%-14% of normal morphology; and group C, >14% of normal morphology). The criteria for the sperm-head vacuoles were those given in the World Health Organization manual. For the analysis of the age factor, the participants were divided into the following groups: 26-30 years, 31-35 years, 36-40 years, 41-45 years, and 46-50 years. Results: The percentage of sperm-head vacuoles increased with normal sperm morphology (group A vs. groups B, C) (p<0.05). In the case of the age factor, a statistically significant difference was not observed across any of the age groups. Conclusion: A majority of the sperm-head vacuoles showed a statistically significant difference among normal morphology groups. Therefore, we should consider the probability of the percentage of sperm-head vacuoles not increasing with age but with abnormal sperm morphology. A further study is required to clarify the effect of the sperm-head vacuoles on ART outcomes.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.2
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• pp.97-101
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• 2016

Objective: Growth hormone and its mediator, insulin-like growth factor-1 (IGF-1), have been suggested to exert gonadotropic actions in both humans and animals. The present study was conducted to assess the relationship between serum IGF-1 concentration, seminal plasma concentration, and sperm parameter abnormalities. Methods: A total of 79 men were enrolled in this study from December 2011 to July 2012 and were prospectively analyzed. Patient parameters analyzed included age, body mass index, smoking status, urological history, and fertility history. Patients were divided into four groups based on their semen parameters: normal (A, n=31), abnormal sperm motility (B, n=12), abnormal sperm morphology (C, n=20), and two or more abnormal parameters (D, n=16). Patient seminal plasma and serum IGF-1 concentrations were determined. Results: Patient baseline characteristics were not significantly different between any of the groups. The serum IGF-1 levels in groups B, C, and D were significantly lower than the levels in group A; however, the seminal plasma IGF-1 levels were not significantly different between any of the groups. Conclusion: Men with abnormal sperm parameters had significantly lower levels of serum IGF-1 compared with men with normal sperm parameters. Seminal plasma IGF-1 levels, however, did not differ significantly between the groups investigated here. Further investigations will be required to determine the exact mechanisms by which growth hormone and IGF-1 affect sperm quality.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.2
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• pp.77-82
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• 2002

Objective : This study is to evaluate the efficacy of intracytoplasmic sperm injection (ICSI) for previous fertilization failure with conventional in vitro fetrtilization (IVF), compared with ICSI for male factor. Method: The author analyzed the 3 years of clinical experience with ICSI retrospectively, between the conventional IVF failure group (IVF failure) and male factor group (male factor). Surgically retrieved epididymal or testicular spermatozoa for ICSI were excluded. The IVF failure group was 13 cycles of 6 patients and male factor group was 30 cycles of 15 patients. Results: The fertilization rates of the IVF failure group and male factor group were 63% and 66% respectively (p=0.635). The clinical pregnancy rates of the both group were 23.1% and 26.7% (p=0.804), and that of live birth rates were 15.4% and 13.3% (p=0.858). There were no significant difference between the two groups. Conclusion: The author concluded that ICSI can overcome previous fertilization failure, with the same fertilization and clinical pregnancy rates seen in patients with male factor.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.1
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• pp.61-68
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• 2021

Objective: This study was conducted to investigate the relationship of semen parameters in samples used for intracytoplasmic sperm injection (ICSI) with fertilization and pregnancy rates in infertile couples. Methods: In this prospective study of Infertile couples with male factor infertility that had undergone ICSI, fractions of the same semen samples obtained for microinjection (to ensure the best predictability) were evaluated to determine the semen parameters and sperm DNA fragmentation index (DFI) on the day of oocyte recovery. Results: In total, 120 couples completed the study and were subdivided into fertilized (n=87) and non-fertilized couples (n=33). The fertilized couples were further classified into pregnant (n=48) and non-pregnant (n=39) couples. Compared to non-fertilized and non-pregnant couples, fertilized and pregnant couples showed statistically significantly higher sperm viability and percentage of normal sperm morphology, as well as significantly lower sperm DFI values. A receiver operating characteristic curve analysis of data from the 120 ICSI cycles showed that sperm viability, normal sperm morphology percentages, and sperm DFI were significant prognostic indicators of fertilization at cutoff values of 40%, 7%, and 46%, respectively. A sperm DFI of 46% showed sensitivity and specificity of 95% and 90%, respectively, for predicting fertilization, and no clinical pregnancies occurred in couples with a sperm DFI above 46%. Conclusion: Semen parameters from the ICSI day sample, especially sperm viability, normal morphology, and DFI, had an impact on fertilization and pregnancy outcomes in ICSI cycles.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.4
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• pp.297-306
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• 2020

The aims of the present study were to confirm that regulation of the PA and environment via TGF-β regulation of sperm by Percoll-separated in porcine uterine epithelial cells. And, it was performed to identify the cytokines (TGF-β1, 2 and 3, TGF-β receptor1 and 2; interleukin, IL-6, IL-8) and PA-related genes (urokinase-PA, uPA; tissue-PA, tPA; PA inhibitor, PAI; uPA-receptor, uPAR) by spermatozoa. The experiment used porcine uterus epithelial cells (pUECs) and uterine tissue epithelial cells, Boar sperm were separated by discontinuous Percoll density gradient (45/90%), and tissues were co-incubated with spermatozoa, followed by real-time PCR. PA activity was measured of sperm by discontinuous Percoll density gradient (45/90%) for 24 hours. To measure viability and acrosome damage of sperm double stained propidium iodide (PI) and SYBR-14 or FITC-PNA were used. In results, binding ratio of Percoll-separated sperm was found no differences, but sperms isolated from 90% Percoll layer reduced PA activity (p < 0.05). when co-cultured sperm selected Percoll in porcine uterus tissues epithelial cells, 90% layer sperm increased TGF-β R1, contrastively tPA and PAI-1 in comparison with control (p < 0.05). 45% sperm was decreased the expression of uPA (p < 0.05). TGF-β decreased PA activity in the supernatant collected from pUECs (p < 0.05). Especially, The group including uPA, PAI-1 were induce sperm intact, while it was reduced in sperm damage when compared to control (p < 0.05). Also, there was no significant difference group of tPA and tPA+I in the dead sperm and acrosome damage compared to control. The expression of tPA and PAI showed a common response. Percoll-separated spermatozoa in 90% layer reduced tPA and IL-related gene mRNA expression. Thus, Percoll-sparated sperm in 90% layer show that it can suppress inflammation through increased expression of TGF-β and downregulation of PA and IL in epithelial cells compared to 45% layer Percoll.

• Development and Reproduction
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• v.21 no.3
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• pp.229-235
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• 2017

In the field of reproductive medicine, assessment of sperm motility is a key factor for achieving successful artificial insemination, in vitro fertilization, or intracellular sperm injection. In this study, the motility of boar sperms was estimated using real-time imaging via confocal microscopy. To confirm this confocal imaging method, flagellar beats and whiplash-like movement angles were compared between fresh and low-temperature-preserved ($17^{\circ}C$ for 24 h) porcine sperms. Low-temperature preservation reduced the number of flagellar beats from $11.0{\pm}2.3beats/s$ (fresh sperm) to $5.7{\pm}1.8beats/s$ and increased the flagellar bending angle from $19.8^{\circ}{\pm}13.8^{\circ}$ (fresh) to $30.6^{\circ}{\pm}15.6^{\circ}$. These data suggest that sperm activity can be assessed using confocal microscopy. The observed motility patterns could be used to develop a sperm evaluation index and automated confocal microscopic sperm motility analysis techniques.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.27-34
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• 1997

This study aimed to evaluate the effect of seminal vesicle fluid (SVF) on the acrosome reaction (AR) occurred spontaneously or induced by $Ca^{2+}$ ionophore A23187, follicular fluid, and progesterone in mouse epididymal sperm. SVF was divided into high (MW>10 kD) and low (MW<10 kD) fractions by ultrafiltration. The low MW fraction of SVF decreased the rate of spontaneous AR, however the high MW fraction did not. It suggested that the low MW fraction of SVF might have contained decapacitation factor(s) responsible for prolonging of time need for capacitation. When sperm preincubated for 60 min in the presence of SVF, the rate of AR induced by A23187 was decreased, but prolongation of preincubation time for 120 min significantly potentiated the AR by A23187. It suggested that addition of SVF into sperm preincubation medium imposed the epididymal sperm a condition similar to ejaculation. AR induced by human follicular fluid or progesterone was also inhibited by SVF. It suggested that substance in SVF might have affected AR of mouse sperm by inhibiting the interaction between AR inducing ligands and sperm surface receptors involved in acrosomal exocytosis.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.3
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• pp.101-105
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• 2015

Objective: Several publications have established a relationship between sperm DNA damage and male factor infertility, based on data from America, Europe, and Asia. This study aimed to compare the extent of sperm DNA damage in sperm samples from Nigerian men with unexplained infertility and in sperm samples from a fertile group composed of sperm donors who had successfully impregnated a female partner naturally or through assisted conception. Methods: A total of 404 men underwent male fertility evaluation at Androcare Laboratories and Cryobank participated in this study. Semen analysis and a sperm chromatin structure assay (SCSA) were performed on all subjects. Results: The men in the unexplained infertility group were slightly older than the men in the fertile sperm group ($36{\pm}10$ years vs. $32{\pm}6$ years, p=0.051). No significant difference was observed between the two groups in semen analysis parameters ($p{\geq}0.05$). Men in the unexplained infertility group with normal semen parameters had a significantly higher DNA fragmentation index (DFI) than men in the fertile sperm group ($27.5%{\pm}7.0%$ vs. $14.1%{\pm}5.3%$, p<0.05). In the unexplained infertility group, 63% of the men had a DFI greater than 20%, compared to 4% in the fertile sperm group. In the unexplained infertility group, 15.2% of the subjects had a DFI greater than 30%, compared to 1% in the fertile sperm group. Conclusion: Our study showed that the SCSA may be a more reliable predictor of fertility potential than traditional semen analysis in cases of unexplained infertility.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.2
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• pp.45-50
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• 2015

Objective: Artificial oocyte activation (AOA) is an effective method to avoid total fertilization failure in human in vitro fertilization-embryo transfer (IVF-ET) cycles. AOA performed using a calcium ionophore can induce calcium oscillation in oocytes and initiate the fertilization process. We evaluated the usefulness of AOA with a calcium ionophore in cases of total fertilization failure in previous cycles and in cases of severe male factor infertility patients with non-motile spermatozoa after pentoxifylline (PF) treatment. Methods: The present study describes 29 intracytoplasmic sperm injection (ICSI)-AOA cycles involving male factor infertility at Cheil General Hospital from January 2006 to June 2013. Patients were divided into two groups (control, n=480; AOA, n=29) depending on whether or not AOA using a calcium ionophore (A23187) was performed after testicular sperm extraction-ICSI (TESE-ICSI). The AOA group was further split into subgroups according to sperm motility after PF treatment: i.e., motile sperm-injected (n=12) and non-motile sperm-injected (n=17) groups (total n=29 cycles). Results: The good embryo rate (52.3% vs. 66.9%), pregnancy rate (20.7% vs. 52.1%), and delivery rate (10.3% vs. 40.8%) were lower in the PF/AOA group than in the control group. When evaluating the effects of restoration of sperm motility after PF treatment on clinical outcomes there was no difference in fertilization rate (66.6% vs. 64.7% in non-motile and motile sperm, respectively), pregnancy rate (17.6% vs. 33.3%), or delivery rate (5.9% vs. 16.7%) between the two groups. Conclusion: We suggest that oocyte activation is a useful method to ensure fertilization in TESE-ICSI cycles regardless of restoration of sperm motility after PF treatment. AOA may be useful in selected patients who have a low fertilization rate or total fertilization failure.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.261-266
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• 1994

Semen preparation for Assisted Reproductive Technology(ART) involves the separation of the motile sperm from non-motile, dead sperm, cellular debris and seminal fluid. The aim of this study is to compare the motile sperm recovery rate and motility index of different sperm preparation method(swim-up method, 80% isotonic continuous percoll method, two-layer discontinuous percoll method, mini-percoll method). In Mini-Percoll method, pellets from patients were suspended in 0.3ml of medium and layerd on a discontinuous percoll gradient consisting of 0.3ml each of 50, 70, 95% isotonic percoll. All semen samples are divided into normal and subnormal sperm group(oligo-, astheno-, oligoasthenozoospermia). Especially, we evaluated the effect of mini-percoll method in subnormal sperm group. In normal sperm group, mini-percoll method and two layer discontinuous percoll method (40%/80%) allowed increasing of motile sperm recovery rate. But motility index was higher in swimup method than the other methods. In subnormal sperm group, mini-percoll method has advantages as compare with the other methods in motile sperm recovery rate and motility index. These results suggest that modified mini-percoll method could be certainly a valuable tool in some cases of severe male factor sperm.