• Clinical and Experimental Reproductive Medicine
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• 제24권3호
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• pp.311-318
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• 1997

Bypassing acrosome reaction and fusion process in intracytoplasmic sperm injection(ICSI), most of injected spermatozoa still contain intact acrosome contents and plasma membrane. It Is not known yet what acrosome contents and plasma membrane of spermatozoa have effect on the development of embryo. For further understanding of fertilization process after ICSI, we studied the time of pronucleus formation, disappearance and first cleavage in human zygote, and pregnancy rate in relation to acrosome reaction rate of spermatozoa after ICSI. Seventy cycles undergoing ICSI program were randomly selected. Sperm suspension from 38 cycles were treated 50% human follicular fluid(hFF) for 3 hours in order to induce acrosome reaction, others were not treated as control. Acrosome reaction in hFF treated and non-treated group was assessed by fluorescein isothiocyanate(FITC)-conjugated Arachis hypogea(PNA) and Pisum sativum agglutinin(PSA). Oocytes were classified into 'good' and 'poor' according to their morphology. After ICSI, fertilization of oocytes were assessed by detection of two pronuclei at 16 hours. The pronuclei disappearance and first cleavage of zygotes were observed at 24 hours, and then embryos were transferred to uterus after culture for 72 hours. The rate of acrosome reaction of spermatozoa in hFF treated group was significantly higher than that in control(p<0.01). Fertilization rates of good oocytes were not different both control and hFF treated group(81.3%(174/206) vs. 72.1%(102/130)). But, in poor oocytes, the fertilization rates in hFF treated group(72.1%(149/183)) were increased compared than those of control group (63.6%(98/140), p<0.01). In either good or poor oocytes, the rates of pronuclei disappearance in hFF treated-spermatozoa injected oocytes were higher than control (59.1%(103/174), 56.4%(84/149) vs. 32.4%(33/102), 37.8%(37/98), p<0.01). Also, the rates of thirst cleavage were increased in hFF treated group (31%(54/174), 24.1%(36/149)) compared than those of control group (10.8%(11/102), 13.2%(13/98), p<0.01). The pregnancy rates of hFF treated group (42.1%(16/38)) were slightly higher than control group (28.1%(9/32), p>0.05). But, the pregnancy rate of group which possessed more than one cleavaged zygote at 24 hours was higher than group which did not (45.2%(19/42) vs. 21.4%(6/28), p<0.05). From these results, the development of zygotes were faster in higher acrosome reacted sperm group than lower acrosome reacted sperm group after ICSI. Our results may be explained that acrosomal membrane and plasma membrane are easily detached from spermatozoa in acrosome reacted spermatozoa compared with acrosome intact sperm in the cytoplasm of oocyte during pronuclear formation. We conclude that the injection of acrosome reacted spermatozoa will increase the pregnancy rate as they can induce fast embryonic development in ICSI.

• 한국수정란이식학회지
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• 제17권1호
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• pp.45-53
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• 2002

본 연구는 sperm-mediated gene transfer를 이용하여 ICSI에 의한 형질전환동물 생산의 기초자료로서 활용하기 위해, ICSI에 사용될 정자의 조건과, 그에 따라 적합한 돼지 정자와 외래유전자의 전처리 및 ICSI를 통한 수정을 및 후기 배로의 발달율과 외래유전자의 발현 여부를 조사하여 다음과 같은 결과를 얻었다. ICSI에 이용될 정자의 조건에 따라 정소상체미부정자, 사출정자 및 동결정자를 이용하여 ICSI후 수정율은 각각 72.3%, 64.1% 및 74.1%로 나타나 유의적인 차이를 나타내지 않았으며, 또한 후기배로의 발달율에 있어서도 각각 17.6%, 18.7% 및 15.0%로 나타나 각 처리군간의 유의적인 차이는 나타나지 않았다. 그리고, ICSI후 전기적 자극을 실시한 군과 실시하지 않은 군에서 난자의 활성화에 따른 수정율은 대조구로 이용한 shame injection과 전기적 활성화를 실시하지 않은 군에서 각각 47.1%와 46.3%로 나타나 전기적 활성화를 실시한 군의 79.6%에 비해 유의적인 차이를 나타내었다. 후기배로의 발달율에 있어서도 전기적 활성화를 실시한 군에서는 24.1%로 나타나 전기 적 활성화를 실시하지 않은 군에서의 14.4%와 유의적인 차이를 나타내었다. 그리고 대조구로 이용한 shame injection에 있어서 후기 배로의 발달율은 2.5%로 낮은 결과를 나타내었다. 또한, 정자와 pcDNA LacZ유전자의 처리시 electroporation 방법을 실시하여 ICSI후 난자를 각각 전기적 활성화를 실시한 군과 실시하지 않은 군에 있어서 유전자 발현율은 각각 30.2%와 24.2% 나타났으나, 두 처리군간에 유의적인 차이는 나타나지 않았다. 그러나, 두 군에서 pcDNA LacZ 유전자는 모두 mosaic 발헌 양상을 보였다. 이상의 실험 결과들을 종합해 보면, ICSI에 사용될 수 있는 돼지의 정자는 정소상체미부정자, 사출정자 및 동결정자 모두가 이용 가능하며, ICSI후 추가 전기적 자극에 의한 난자의 활성화가 수정율과 후기 배로의 발달율을 향상시킬 수 있음을 시사하였다. 따라서, 돼지에 있어서 정자의 외래유전자 도입에 대한 정확하고 실용적인 방법은 보고되고 있지는 않은 상태로, 정자와 외래유전자의 처리법을 향상시키기 위하여 다양한 방법으로 많은 연구가 요구된다.

• 한국수정란이식학회지
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• 제24권3호
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• pp.145-150
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• 2009

This study was conducted to examine the effects of human follicular fluid and gonadotropin (FSH+HCG+rhEGF) on in vitro maturation, fertilization and development of human immature oocytes. Cumulus-oocyte complexes (COCs) were collected following for in vitro fertilization and embryo transfer (IVF-ET) cycles of the patients. At the time of oocytes collection, oocytes were classified into MII, MI and GV in accordance with their appearance (MII: Fully mature oocyte at metaphase II of meiosis; MI: Nearly mature oocytes at metaphase I of meiosis; GV: Immature oocytes at prophase I of meiosis). After controlled ovarian stimulation using gonadotropin(FSH) and human chorionic gonadotropin (HCG) in 70 ICSI cycles, 158 MI to MII matured oocytes were intracytoplasmic sperm injection (ICSI) ${\sim}4$ h after in vitro culture and 553 MII oocytes were ICSI after denudation. The aspirated MI and GV oocytes were cultured in culture medium containing 10% (v/v) serum protein substitute (SPS), 10% (v/v) human follicular fluid (hFF) and 10% (v/v) serum protein substitute (SPS)+1 IU/ml FSH+10 IU/ml HCG+10 ng/ml recombinant human epidermal growth factor (rhEGF). The maturation rate of immature oocytes was similar among the three group. When maturation medium was supplemented with 10% SPS, 10% hFF or gonadotropins, the fertilization rate of in vitro matured oocytes was higher in 10% SPS (80.0%), but there was no statistical significance (78.2%; hFF, 76.9%; gonadotropin, p>0.05). The development rate of human embryos developed to $6{\sim}8$ cells were not significant difference in the medium containing SPS, hFF and gonadotropins (65.6%, 65.9% and 66.7%). The results of these study suggest that human follicular fluid and gonadotropins supplemented in the culture medium was not effected on the in vitro maturation, fertilization and development of human immature oocytes.

• Clinical and Experimental Reproductive Medicine
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• 제43권1호
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• pp.38-43
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• 2016

Objective: The number of assisted reproductive technology (ART) clinics, ART cycles, clinical pregnancy rate (CPR), and number of newborns conceived using ART have steadily increased in South Korea. This aim of this study was to describe the status of ART in South Korea between January 1 and December 31, 2011. Methods: A localized online survey was created and sent to all available ART centers via email in 2015. Fresh embryo transfer (FET) cases were categorized depending on whether standard in vitro fertilization, intracytoplasmic sperm injection (ICSI), or half-ICSI procedures were used. Thawed embryo transfer (TET) and other related procedures were surveyed. Results: Data from 36,990 ART procedures were provided by 74 clinics. Of the 30,410 cycles in which oocytes were retrieved, a complete transfer was performed in 91.0% (n=27,683). In addition, 9,197 cycles were confirmed to be clinical pregnancies in the FET cycles, representing a pregnancy rate of 30.2% per oocyte pick-up and 33.2% per ET. The most common number of embryos transferred in the FET procedures was three (38.1%), followed by two (34.7%) and one (14.3%). Of the 8,826 TET cycles, 3,137 clinical pregnancies (31.1%) were confirmed by ultrasonography. Conclusion: While the overall clinical pregnancy rate for the TET cycles performed was lower than the rate reported in 2010 (31.1% vs. 35.4%), the overall CPR for the FET cycles was higher than in 2010 (33.2% in 2011 and 32.9% in 2010). The most common number of embryos transferred in FET cycles was three, as was the case in 2010.

• 대한간호학회지
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• 제50권3호
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• pp.369-384
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• 2020

Purpose: The purpose of this study was to analyze anxiety and depression among infertile women at different time points during the first In Vitro Fertilization (IVF) or Intracytoplasmic Sperm Injection (ICSI) treatment through a systematic review and meta-analysis. Methods: Seven out of 3,011 studies were included for meta-analysis. To estimate the effect size, a meta-analysis of the studies was performed using the RevMan 5.3 program. We compared the measurement outcomes at three time points: before the start of treatment (T0), cancellation of treatment after pregnancy detection (T2), one to six months after treatment (T3). The effect size used was the standardized mean difference (SMD). Results: In comparing the different time points of the pregnant women from their cycle, significantly lower levels of depression were found at T2 than at T0. In non-pregnant women, anxiety at T2 and depression at T2 and T3 were significantly higher than those at T0. At T2 and T3, the non-pregnant women reported higher levels of anxiety and depression compared with the pregnant women. Conclusion: Anxiety and depression in infertile women undergoing the first IVF or ICSI are associated with the time points and pregnancy status after treatment. These findings suggest that attention should be paid to helping infertile women prepare for and cope with treatment and treatment failure.

• Clinical and Experimental Reproductive Medicine
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• 제44권4호
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• pp.224-231
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• 2017

Objective: We studied the association between sperm DNA fragmentation (SDF) and several clinical in vitro fertilization outcomes. Methods: We retrospectively analyzed 169 consecutive fresh IVF cycles. Semen was collected on the day of oocyte retrieval, and we assessed standard semen parameters and the SDF level (by terminal deoxynucleotidyl transferase dUTP nick-end labeling). Poor ovarian response (POR) was defined as the collection of three or fewer mature oocytes. Oocytes were inseminated by the conventional method or intracytoplasmic sperm injection. Results: SDF did not affect the fertilization or pregnancy rate, but did have a significant effect on the miscarriage rate. In the miscarriage group (n = 10), the SDF level was significantly higher (23.9% vs. 14.1%) and number of mature oocytes was significantly lower (4.3 vs. 7.6) than in the live birth group (n = 45). Multiple regression analysis showed that SDF was an independent predictor of miscarriage (odds ratio, 1.051; 95% confidence interval, 1.001-1.104). The cutoffs for the SDF level and number of mature oocytes that could predict miscarriage were > 13% and ${\leq}3$, respectively. In the low-SDF group (${\leq}13%$), the miscarriage rate was similar in POR patients and those with a normal ovarian response (NOR; 14.2% vs. 4.3%). In the high-SDF group ( > 13%), the miscarriage rate was significantly higher in the POR group than in the NOR group (60.0% vs. 13.3%, p= 0.045). Conclusion: Our study demonstrated that a high SDF level ( > 13%) was associated with a high miscarriage rate, and that it mainly contributed to miscarriage in the POR group. The results suggest that SDF measurements should be considered in couples with POR in order to predict the prognosis of the pregnancy.

• Clinical and Experimental Reproductive Medicine
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• 제44권4호
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• pp.232-238
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• 2017

Objective: To determine whether reducing the cetrorelix dose in the antagonist protocol to 0.125 mg had any deleterious effects on follicular development, the number and quality of retrieved oocytes, or the number of embryos, and to characterize its effects on the affordability of assisted reproductive technology. Methods: This randomized controlled study was conducted at the Fertility Unit of Tanta Educational Hospital of Tanta University, the Egyptian Consultants' Fertility Center, and the Qurrat Aien Fertility Center, from January 1 to June 30, 2017. Patients' demographic data, stimulation protocol, costs, pregnancy rate, and complications were recorded. Patients were randomly allocated into two groups: group I (n = 61) received 0.125 mg of cetrorelix (the study group), and group II (n = 62) received 0.25 mg of cetrorelix (the control group). Results: The demographic data were comparable regarding age, parity, duration of infertility, and body mass index. The dose of recombinant follicle-stimulating hormone units required was $2,350.43{\pm}150.76$ IU in group I and $2,366.25{\pm}140.34$ IU in group II, which was not a significant difference (p= 0.548). The duration of stimulation, number of retrieved oocytes, and number of developed embryos were not significantly different between the groups. The clinical and ongoing pregnancy rates likewise did not significantly differ. The cost of intracytoplasmic sperm injection per cycle was significantly lower in group I than in group II (US $ $494.66{\pm}4.079$ vs. US $ $649.677{\pm}43.637$). Conclusion: Reduction of the cetrorelix dose in the antagonist protocol was not associated with any significant difference either in the number of oocytes retrieved or in the pregnancy rate. Moreover, it was more economically feasible for patients in a low-resource country.

• 대한의생명과학회지
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• 제10권1호
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• pp.31-34
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• 2004

White blood cells (WBCs) are present in most human ejaculates, but abnormally high concentration of seminal leukocytes may reflect an underlying pathological condition. The World Health Organization (WHO) has defined leukocytospermia as status of more than $10^6$ WBC/mL of semen. The purpose of this study was firstly, to compare the outcomes between conventional IVF and ICSI in leukocytospermia, and secondly, to investigate whether ICSI may be an alternation treatment for patients with leukocytospermia. Total 346 cycles of conventional IVF and ICSI candidates underwent IVF cycles at Eulji Hospital Infertility Clinic. Semen Parameters including concentration, motility, morphology of spermatozoa and concentration of leukocytes were assessed from the raw ejaculates. There was no difference in sperm concentration, motility and morphology. The rates of fertilization and good embryo development from ICSI were significantly higher than those from conventional IVF in leukocytospermia (60.4% ＆ 32.5%, respectively for ICSI group and 44.4% ＆ 28.5%, respectively for IVF group, P<0.00l). The pregnancy rate after ICSI was also higher than that from conventional IVF (34.0% vs 29.1 %, P<0.05). These results indicate that the presence of seminal leukocyes (> 1$\times10^6$ WBC/mL of semen) is adversely related with fertilization, embryo development and pregnancy rate. Therfore the measurement of seminal leukocytes in routine semen analysis appears to be of prognostic value with regard to male fertilizing potential. In conclusion, it is suggested that ICSI is an alternative choice of treatment for patients with leukocytospermia.

• Journal of Preventive Medicine and Public Health
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• 제50권6호
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• pp.347-360
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• 2017

Objectives: This meta-analysis aimed to evaluate congenital malformations in infants conceived by assisted reproductive techniques (ART), compared with infants conceived spontaneously. Methods: In this study, available resources searched to find relevant articles included PubMed, ScienceDirect, Scopus, Google Scholar, Cochrane, ProQuest, Iranmedex, Magiran, and Scientific Information Database. After extracting the necessary information from evaluated articles, meta-analysis on the articles' data was performed using Stata version 11.2. Results: In this study, from a total of 339 articles, extracted from the initial investigation, ultimately 30 articles were selected for meta-analysis that assessed the use of ART on the risk of congenital abnormalities and some birth complications on 5 470 181 infants (315 402 cases and 5 154 779 controls). The odds ratio (95% confidence interval [CI]) for low birth weight was 1.89 (95% CI, 1.36 to 2.62), preterm labor 1.79 (95% CI, 1.21 to 2.63), cardiac abnormalities 1.43 (95% CI, 1.27 to 1.62), central nervous system abnormalities 1.36 (95% CI, 1.10 to 1.70), urogenital system abnormalities 1.58 (95% CI, 1.28 to 1.94), musculoskeletal disorders 1.35 (95% CI, 1.12 to 1.64), and chromosomal abnormalities in infants conceived by ART was 1.14 (95% CI, 0.90 to 1.44), which were all statistically significant, except chromosomal abnormalities. Conclusions: The risk of congenital abnormalities and some birth complications were significantly higher in ART than normal conception, while chromosomal abnormalities were not; therefore, the application of ART should be selected individually for patients by detailed assessment to reduce such risks in the population.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.26-26
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• 2003

Hyaluronic acid (HA)-binding sites have been shown the diagnostic potential fur assessment of sperm maturity, which is related to male fertility. This study was designed to evaluate chromosomal patterns in porcine embryos produced by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with non- or HA-binding sperm (HABS). For binding of sperm with HA, sperm incubated in 10 ${mu}ell$ drop containing HA (0.8 mg/ml)-agarose (0.8%) mixture for 15 min. IVF and ICSI with non- or HA-bound sperm examined with matured oocytes at 44 hr after in vitro maturation. Embryos were cultured in 50 ${mu}ell$ of NCSU 23 containing 0.5% BSA for 5 days and then in 50 ${mu}ell$ of NCSU 23 containing 10% FBS for 2 days. For the evaluation of chromosomal aneuploidies, chromosome 1 sub-metacentric specific probe was used in sperm and embryos by fluorescence in situ hybridization (FISH). The frequency of aneuploidy sperm for chromosome 1 was 6.25%. The significant differences following IVF and ICSI with non- or HA-bound sperm were not observed in blastocyst formation rates (18.6, 23.5, and 23.8%) and cell number (61.8 $\pm$ 12.5, 55.5 $\pm$ 7.3, and 59.3 $\pm$ 9.6). Moreover, the percentage of diploidy in 4-cell stage embryos was 57.1% (IVF), 68.8% (ICSI), and 76.3% (ICSI-HABS). These results suggest that HA-binding sites may be a material for selection of normal sperm for ICSI. Therefore HA selection of normal sperm may be reduce the loss to embryonic mortality prior to embryo transfer in pig.