Kim, Eun-Kyung;Kim, Eun-Ha;Kim, Eun-Ah;Lee, Kyung-Ah;Shin, Ji-Eun;Kwon, Hwang
Clinical and Experimental Reproductive Medicine
/
v.42
no.1
/
pp.22-29
/
2015
Objective: Sperm must be properly prepared in in vitro fertilization (IVF)-embryo transfer (ET) programs in order to control the fertilization rate and ensure that embryos are of high quality and have appropriate developmental abilities. The objective of this study was to determine the most optimal sperm preparation method for IVF. Methods: Patients less than 40 years of age who participated in a fresh IVF-ET cycle from November 2012 to March 2013 were included in this study. Poor responders with less than three mature oocytes were excluded. Ham's F-10 medium or sperm-washing medium (SWM) was used in combination with the density-gradient centrifugation/swim-up (DGC-SUP) or SUP methods for sperm preparation. A total of 429 fresh IVF-ET cycles were grouped according to the media and methods used for sperm preparation and retrospectively analyzed (DGC-SUP/Ham's F-10, n=82; DGC-SUP/SWM, n=43; SUP/Ham's F-10, n=181; SUP/SWM, n=123). Results: There were no significant differences among these four groups with respect to the mean age of the female partners, duration of infertility, number of previous IVF cycles, and retrieved oocytes. We determined that both the DGC-SUP and SUP methods for sperm preparation from whole semen, using either Ham's F-10 or SWM media, result in comparable clinical outcomes, including fertilization and pregnancy rates. Conclusion: We suggest that both media and both methods for sperm preparation can be used for selecting high-quality sperm for assistive reproductive technology programs.
Septin4 belong to a family of polymerizing GTP-binding proteins that are required for many cellular functions, such as membrane compartmentalization, vesicular trafficking, mitosis, and cytoskeletal remodeling. Since, Septin4 is expressed specifically in the testis, we aimed to determine the association between Septin4 gene expression with sperm quality, DNA damage, and stress oxidative level in infertile patients. The present study included 60 semen samples that grouped into three groups: normozoospermia (n=20), asthenozoospermia (n=20), astheno-teratozoospermia (n=20). Initially, semen parameters were analyzed by using the World Health Organization protocol. The mRNA expression of Septin4 in sperm was examined using reverse transcription-polymerase chain reaction. Oxidative stress markers, i.e., total antioxidant capacity, superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde, were determined by ELISA kit. The current study showed a statistically significant highly positive correlation in Septin4 gene expression with sperm motility, normal morphology, viability, capacity, and sperm mitochondrial membrane potential (MMP). However, it showed significant negative correlation with sperm DNA fragmentation. Septin4 had a significant correlation with stress oxidative factor and antioxidant enzyme levels. In conclusion, Septin4 gene expression provides clinical useful information for the diagnosis of male infertility. It might be a marker for discrimination between fertile and infertile patients. The current study showed a statistically significant highly positive correlation in Septin4 gene expression with sperm motility, normal morphology, viability, capacity, and sperm MMP. However, it shows significant negative correlation with sperm DNA fragmentation. Septin4 had a significant correlation with stress oxidative factor and antioxidant enzyme levels.
In a summer study during May to July, involving 12 young Murrah buffalo bulls at forty months of average age, the effects of multiple shower vs single shower body cooling and vitamin A, D and E supplementation on the sexual behaviour, semen quality and freezability were investigated. The animals were divided into two groups (6 animals in each group) and housed in a half-walled shed with proper spacing, the feeding management being identical. The bulls in the control group were given a single shower at 1000 h, whereas the experimental bulls were given four showers at 10,12,14 and 16 h. In addition, the experimental bulls were given vitamin A, D and E injections at fifteen day intervals. The sexual behaviour of bulls was observed in terms of reaction time, sexual aggressiveness and ejaculatory thrust. Semen quality of all the bulls was assessed in terms of volume, mass activity, live-dead sperm and sperm concentration, sperm motility and morphology, and acrosomal abnormality. The sexual behaviour did not vary significantly between the groups, whereas semen quality differed significantly for volume, per cent live sperms, total sperms per ejaculate and total live sperm per ejaculate between groups. It can be concluded that sexual behaviour was not influenced by the thermal comfort treatment coupled with periodic vitamin A, D and E injections. But the treatments improved most of the seminal traits in the experimental group of bulls. However, benefit of treatment was not reflected in the freezability traits of the semen.
Choi, Hye Won;Park, Yong-Seog;Lee, Sun-Hee;Lim, Chun Kyu;Seo, Ju Tae;Yang, Kwang Moon
Clinical and Experimental Reproductive Medicine
/
v.43
no.4
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pp.221-227
/
2016
Objective: The aim of this study was to evaluate the influence of maternal age on fertilization, embryo quality, and clinical pregnancy in patients undergoing intracytoplasmic sperm injection (ICSI) using testicular sperm from partners with azoospermia. Methods: A total of 416 ICSI cycles using testicular spermatozoa from partners with obstructive azoospermia (OA, n = 301) and non-obstructive azoospermia (NOA, n = 115) were analyzed. Female patients were divided into the following age groups: 27 to 31 years, 32 to 36 years, and 37 to 41 years. The rates of fertilization, high-quality embryos, clinical pregnancy, and delivery were compared across maternal age groups between the OA and NOA groups. Results: The rates of fertilization and high-quality embryos were not significantly different among the maternal age groups. Similarly, the clinical pregnancy and delivery rates were not significantly different. The fertilization rate was significantly higher in the OA group than in the NOA group (p< 0.05). Age-group analysis revealed that the fertilization and high-quality embryo rates were significantly different between the OA and NOA groups in patients aged 27 to 31 years old, but not for the other age groups. Although the clinical pregnancy and delivery rates differed between the OA and NOA groups across all age groups, significant differences were not observed. Conclusion: In couples using testicular sperm from male partners with azoospermia, pregnancy and delivery outcomes were not affected by maternal age. However, women older than 37 years using testicular sperm from partners with azoospermia should be advised of the increased incidence of pregnancy failure.
Ma, Lei;Jung, Dae-Jin;Jung, Eun-Ju;Lee, Woo-Jin;Hwang, Ju-Mi;Bae, Jeong-Won;Kim, Dae-Hyun;Yi, Jun Koo;Lee, Sang Moo;Ha, Jae Jung;Kwon, Woo-Sung
Journal of Animal Reproduction and Biotechnology
/
v.36
no.2
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pp.106-115
/
2021
Sperm cryopreservation is an important method of assisted reproductive techniques and storing genetic resources. It plays a vital role in genetic improvement, livestock industrial preservation of endangered species, and clinical practice. Consequently, the cryopreservation technique is well organized through various studies, especially on Korean native cattle (Hanwoo). However, the cryopreservation technique of Korean native brindled cattle, which is one of the native cattle species in Korea, is not well organized. Therefore, it is necessary to develop a Supplementary Table technique for the cryopreservation of Korean native brindled cattle. For this purpose, it is important to first evaluate the quality of the currently produced cryopreserved sperm of Korean native brindled cattle. In this study, we randomly selected 72 individual Korean native brindled cattle semen samples collected from 8 different region research centers and used them to evaluate sperm functions. We focused on the quality evaluation of cryopreserved Korean native brindled cattle semen following the measurement of motion kinematics, capacitation status, intracellular ATP level, sperm motility, and cell viability. Then, the values of each of the eight groups were derived from various sperm parameters of nine individual samples, including sperm motility, kinematics, cellular motility, and intracellular ATP levels, which were used to compare and evaluate sperm function. Overall, differences in various sperm parameters were observed between most of the research centers. Particularly, the deviations of motility and motion kinematics were high according to the sample. Therefore, we suggest that it is necessary to develop a standard method for the cryopreservation of Korean native brindled cattle semen. We also suggest the need for sperm quality evaluation of the cryopreserved semen of Korean native brindled cattle before using artificial insemination to attain a high fertility rate.
To maintain a good sperm motility is one of the key factors for the successful artificial insemination in retrograde ejaculation, and the sperm motility has been shown to be affected by various environmental factors, including change in pH and osmolarity. Herein we have analyzed the effect of change in pH and osmolarity in urine and normal saline on sperm motility by Sperm Quality Analyzer and Makler counting chamber. Semen, which sampled by masturbation from a 28 year old male and showed normal finding on semen analysis, was used for this study. The results were as follows: 1. When osmolarity was fixed to 300mOsm, pH did not show a definite effect on the sperm moility. However, the motility was generally a bit better in alkaline urine and saline than in acid, particularly than in pH 5.0. 2. When pH was fixed to 7.5, sperm motility was best in urine and saline of 300mOsm. Hyperosmolarity had more adverse.effect on the motility than hypoosmolarity. 3. The sperm motility was worse in the urine than in saline under the same pH and osmolarity. In conclusion, osmolarity has a definite effect on sperm motility, where as pH has relatively little effect. And certain components of urine other than pH and osmolarity might affect the sperm motility.
Objective: Semen cryopreservation is an effective method of preserving genetic material, particularly in native chicken breeds facing a substantial decline. In this study, we evaluated the quality of frozen/thawed rooster semen treated with different concentrations of oral administrations of black ginger (Kaempferia parviflora: KP) extract and determined its fertility. Methods: Thirty-two Thai native roosters (Pradu Hang Dum, 42 weeks old) were used in this study. The treatments were classified into four groups according to the concentration of KP extract administered to the roosters: 0, 100, 150, and 200 mg/kg body weight. The quality of fresh semen was analyzed before cryopreservation. Post-thaw sperm quality and fertility potential were determined. Also, lipid peroxidation was determined. Results: The results showed that sperm concentration and movement increased in roosters treated with 200 mg/kg of KP extract (p<0.05). The malondialdehyde (MDA) in the roosters receiving 200 mg/kg KP extract was lower than that in the other but had an insignificant difference within the KP treatment groups (p>0.05). The highest MDA levels were observed in the control group (p<0.05). The percentage of motile sperm (total motility and progressive motility) after semen thawing was higher in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (p<0.05). MDA levels decreased significantly in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (p<0.05). Fertility and hatchability were greater in the KP150 and KP200 groups than in the KP100 and control groups (p<0.05). Conclusion: The optimal amount of KP extract influencing initial sperm quality was determined to be 200 mg/kg. However, 150 mg/kg was the optimal low dosage of KP extract administration that maintained sperm quality and fertility following semen cryopreservation.
Park, C.S.;Kim, M.Y.;Yi, Y.J.;Chang, Y.J.;Lee, S.H.;Lee, J.J.;Kim, M.C.;Jin, D.I.
Asian-Australasian Journal of Animal Sciences
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v.17
no.10
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pp.1369-1373
/
2004
The percentages of sperm motility and normal acrosome on the liquid boar semen diluted and preserved at $4^{\circ}C$ with lactose hydrate, egg yolk and N-acetyl-D-glucosamine (LEN) diluent were significant differences according to preservation day and incubation time, respectively. The sperm motility steadily declined from 96.9% at 0.5 h incubation to 78.8% at 6 h incubation at 1 day of preservation. However, the sperm motility rapidly declined after 4 day of preservation during incubation. The normal acrosome steadily declined from 93.3% at 0.5 h incubation to 73.8% at 6 h incubation at 1 day of preservation. However, the normal acrosome rapidly declined after 3 day of preservation during incubation. The rates of sperm penetration and polyspermy were higher in 5 and $10{\times}10^6$ sperm/ml than in 0.2 and $1{\times}10^6$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in $10{\times}10^6$ sperm/ml compared with other sperm concentrations. The rates of blastocysts from the cleaved oocytes (2-4 cell stage) were highest in $1{\times}10^6$sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at $4^{\circ}C$ could be used for in vitro fertilization of pig oocytes matured in vitro. Also, we recommend $1{\times}10^6$sperm/ml concentration for in vitro fertilization of pig oocytes.
The occurrence and time course of capacitation, acrosomal loss, and hyperactivated motility require quantitative definition in order to characterize fertile human sperm. Recently the method has been developed to estimate the quality of spermatozoa by using kinematic parameters such as curvilinear velocity(VCL), average path velocity(VAP), linearity(LIN), straightness(STR), amplitude of lateral head displacement(ALH), and beat cross frequence(BCF) from Computer Assisted Sperm Analysis (CASA). In this study, using the Hamilton Thorn motility analyzer HTM 2030(Hamilton Thorn Research, Beverly, MA), we attempted to identify the spermatozoa with hyperactivated motility (HA) objectively and to monitor hyperactivation of human spermatozoa during incubation in capacitating media and after treatment of calcium ionophore as compared with acrosome status. And we examined whether HA are related to the result of SPA. Semen samples obtained from 16 healthy men were prepared by swim up technique and preincubated in a capacitating media(modified BWW medium) for 5 hours and treated with calcium ionophore solution. The acrosome reaction was detected with PSA-FITC labelling of the acrosome and in vitro sperm ferilizing capacity was assessed by the zona free hamster ovum penetration assay (SPA). The incidence of hyperactivated sperm was 2.6% in fresh semen, 14.3% of the swim up population, 13.7% after 5h of incubation. Significant increase of percentage of hyperactivated sperm was observed after the incubation (p<0.05) but after treatment, no significant changes of percentage of hyperactivated sperm(l1.8%) in contrast to significant rise in the percentage of acrosome reacted cells. Correlation analysis failed to show any significant relationship between the percentage of sperm with HA and SPA score. In conclusion, although no direct correlations were found between the results of SPA and HA, hyperactivation of sperm is associated with capacitation and monitoring hyperactivated sperm will be expected as a method of evaluating the functional quality of sperm such as SPA.
In IVF-ET program, intracytoplasmic sperm injection(ICSI) has been performed with testicular sperm extraction(TESE) in case of no normal spermatozoon could be retrieved from the epididymis. We wished to see whether the quality of testicular sperm affect the fertilization and pregnancy rate in TESE-ICSI cycles(n=40). These cycles were classified into three groups by the total number of normal motile spermatozoa(TNMS) in the TESE sample: i) good sperm(GS) group(n=12), TNMS > 10,000; ii) moderate sperm(MS) group(n=19), 1,000 < TNMS < 10,000; iii) poor sperm(PS) group(n=9), TNMS < 1,000. Among 423 injected oocytes, 307(72.6%) oocytes were normally fertilized and 43 zygotes were cryopreserved. The fertilization rates of GS group(79.3%) and MS group(75.9%) were significantly(p<0.005) higher than PS group(60.2%). After the embryo transfer(n=40), clinical pregnancy was obtained in 14 cycles(35.0%) and on-going pregnacy in 13 cycles(32.5%). The clinical and on-going pregnancy rates were similar in each group. From these results it can be concluded that testicular spermatozoa are successfully used with ICSI in IVF-ET program in spite of very poor quality of TESE sample.
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