• Clinical and Experimental Reproductive Medicine
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• v.50 no.4
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• pp.270-276
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• 2023

Objective: This study investigated the clinical and laboratory factors associated with the presence of dysmorphic oocytes in intracytoplasmic sperm injection (ICSI) cycles. Methods: The study involved 200 ICSI cycles, performed from 2020 to 2021, that yielded at least one mature oocyte. Clinical characteristics and ovarian stimulation methods were compared between 68 cycles with at least one dysmorphic oocyte (the dysmorphic group) and 132 cycles with normal-form oocytes only (the non-dysmorphic group). Dysmorphic oocytes were characterized by dark cytoplasm, cytoplasmic granularity, cytoplasmic vacuoles, refractile bodies in the cytoplasm, smooth endoplasmic reticulum in the cytoplasm, an oval shape, an abnormal zona pellucida, a large perivitelline space, debris in the perivitelline space, or an abnormal polar body. Results: The ages of the women, indications for in vitro fertilization, serum anti-Müllerian hormone levels, and rates of current ovarian endometrioma were similar between the dysmorphic and non-dysmorphic groups. In both groups, the three ovarian stimulation regimens, two types of pituitary suppression, and total gonadotropin dose were employed similarly. However, the dual-trigger method was used more frequently in the dysmorphic group (67.6% vs. 50%, p=0.024). The dysmorphic group contained significantly more immature oocytes and exhibited significantly lower oocyte maturity (50% vs. 66.7%, p=0.001) than the non-dysmorphic cycles. Within the dysmorphic group, significantly lower oocyte maturity was found in the cycles using a dual-trigger, but not in those with a human chorionic gonadotropin trigger. Conclusion: ICSI cycles with dysmorphic oocytes are closely associated with reduced oocyte maturity. This association was observed exclusively in dual-trigger cycles.

• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.339-347
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• 2001

Sperm-mediated DNA transfer has a potential to markedly simplify techniques for the generation of transgenic animals. The exogenous DNA transfer by intracytoplasmic sperm injection (ICSI) procedure has been recently introduced in the production of transgenic animals. In this study, the developmental competence and tile expression rates of transgene were investigated after injection of spermatozoon or sperm head with enhanced green fluorescent protein (EGFP) gene into the mature porcine oocytes. The porcine oocytes were injected with intact sperm, membrane-disrupted sperm or sperm head. After injection. embryos were cultured in NCSU23 medium up to the blastocyst stage, and the developmental competence and expression rates were studied. The developmental rate (67.0%) of sperm injection group was higher than that (59.7%) of sperm head injection group, and the rates of EGFP expression were also significantly different between sperm injection and sperm head injection groups (42.1 vs 20.0%) (F＜0.05). In the porcine oocytes injected with sperm treated with different methods of membrane disruption, the removal of sperm membrane did not alter the developmental competence of embryos. The rate of blastocysts at 7 days after injection with intact and membrane disrupted sperm were 15.0 and 14.2%, respectively. The EGFP expression rates, 38.4% in embryos injected with frozen-thawed sperm was higher than that, 22.4% of embryos injected with the Triton X-100 treated sperm. Prior to injection, sperm were cultured in different EGFP gene concentrations from 0.Ol to 1ng/u${mu}ell$. However, no significant difference in developmental rates of embryos among different concentrations of EGFP gene were observed. The highest expression rate of EGFP gene, 37.4% was obtained from the embryos injected with spermatozoa treated with 0.1 ng/${mu}ell$ EGFP gene. These results suggested that exogenous DNA could be attached to the membrane disrupted sperm, and that these sperm could be used as a vector carrying foreign DNA into embryos.

• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.113-118
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• 1999

This study was carried out to investigate on the improvement of fertilizing ability of in vitro matured oocytes from sperm density and motility by intracytoplasmic sperm injection(ICSI) into the porcine oocytes. 1. The in vitro fertilization and cleavage rates of oocytes from 1.0, 2.0, 3.0, 5.0 ($\times$10$^{6}$ $m\ell$) sperm concentration by IVF and ICSI of porcine oocytes were 46.7%~75.0%, 60.0%~85.7% and 10.6%~25.0%, 20.0%~64.3%, respectively. 2. The in vitro fertilization and cleavage rates of oocytes from 20, 40, 60, 80% of sperm mortilty by IVF and ICSI of porcine oocytes were 46.4%~71.4%, 67.9%~85.7% and 7.1%~21.4%, 28.6%~60.7%, respectively. 3. The in vitro fertilization and developmental rates of oocytes by IVF and ICSI methods were 55.6%~60.0%, 77.8%~80.0% and 17.8%~24.0%, 42.2%~56.0%, respectively. This ICSI method was improved high fertilization rates of porcine oocytes.

• Clinical and Experimental Reproductive Medicine
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• v.22 no.2
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• pp.143-148
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• 1995

To present and assess the efficacy of combination of microsurgical epididymal sperm aspiration(MESA) and intracytoplasmic sperm injection(ICSI) for the treatment of infertility due to unreconstructable obstructive azoospermia or congenital bilateral agenesis of vas deferens (CBAVD), MESA was performed in the 45 husbands ( 16 CBAVD, 29 unreconstructable genital tract obstruction), followed by ICSI of oocytes recovered from the wives hyperstimulated by GnRH agonist in combination with hMG and FSH. Cleaving embryos were transfered to the uterine cavity or follopian tube(ZIFT) 18 or 24 hours after ICSI procedure. In 45 cycles of MESA, 492 oocyte complexes were recovered. ICSI was carried out on 355 metaphase II oocytes and 226 oocytes (63.7%) showed normal two pronucleus fertilization. After 198 embryos were transferred in 43 cycles, an average of 5 per cycle, 20 patients presented a positive HCG and intrauterine pregnancy was confirmed by US. So, the clinical ongoing pregnancy rate per transfer was 46.5%. Until now, 8 patients have given birth to 9 babies, 5 male and 4 female, including 1 twin. The babies were all healthy except 1 twin female baby. There was 1 miscarriage at 7 weeks and chromosomal study of abortus revealed as 45X, monosomy. These results suggested that it was possible to achieve high normal fertilization and pregnancy rate by ICSI using epididymal sperm.

• 대한생식의학회:학술대회논문집
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• 2006.06a
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• pp.66-68
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• 2006

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.181-189
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• 2001

The objective of this study was to evaluate the development of porcine follicular oocytes fertilized by intracytoplasmic sperm injection (ICSI). Cumulus-oocyte-complexes (COCs) were collected by aspiration from follicles of 2~7 mm in diameter from a local slaughterhouse. Oocytes were matured in vitro for 40~44 h, and spermatozoa were prepared by swim-up in the presence or absence of 5 mM dithiothreitol (DTT) and then M II stages of the oocyte were either centrifuged or not centrifuged for the following injection of ooplasm. Injected oocytes were cultured in NCSU 23 medium for 6 to 8 days. The results obtained were as follows: 1. The rates of cleavage and development rates into blastocyst by ICSI were not significantly different between the with (53.0% and 19.7%) or without (48.3% and 23.8%) centrifugation, respectively (P＜0.05). 2. The cleavage and developmental rates to blastocyst after ICSI with or without 5 mM DTT treated-sperm were not significantly different (60.4% vs 16.4% and 45.5% vs 22.2%), respectively (P＜0.05). 3. The cleavage and the developmental rates to btastocyst were not significantly different between the zygotes obtained by IVF (51.8% vs. 22.4%) and ICSI (51.4% vs. 21.6%) (P＜0.05). 4. The number of blastomere in blastocyst stages after IVF or ICSI was not significantly different (46.7$\pm$2.9 and 41.9$\pm$4.6).

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.56-56
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• 2001

The exogenous gene transfer by intracytoplasmic sperm injection (ICSI) procedure has been recently used to produce transgenic mice and pigs. Sperm-mediated DNA transfer has the potential to markedly simplify the generation of transgenic animals. This method may serve as an alternative to the pronucleus injection of DNA for the production of transgenic pigs. Therefore, in this study, we investigated the expression of transgene after co-injection of spermatozoon or sperm head with green fluorescent protein (GFP) gene into in vitro matured porcine oocytes. Spermatozoon and sperm head, that was obtained by sonication, were treated with 0.03% Triton X-100 to remove the membrane. They were preincubated with linearized pEGFP-N1 for 1 min, and then embryos cultured NCSU23 medium for 2.5 days after co-injected of sperm and DNA. We monitored expression of GFP in embryos under epifluorescent microscope. The remove of sperm membrane did not alter the developmental competence of embryos after ICSI. At 7 days following injection, the rates of blastocysts following injection of intact sperm (15.0%), and of sperm with disrupted membrane (14.2%) were higher than that following IVF (10.0%). Porcine oocytes injected with sperm which co-cultured with DNA concentration of 1, 0.1, and 0.01 ng were 60, 65.7 and 75% and 18.5, 37.4 and 22.2% for rates of cleavage and GFP expression, respectively. In vitro matured porcine oocytes injected with sperm and isolated sperm head resulted in 69 and 59.7% of cleavage rates, respectively The rates of embryo GFP expressed did not significantly different between sperm (20.4%) and sperm head (20.0%) injection. The transgenic embryos with the clusters of positive blastomeres were observed under fluorescent microscope. Most of embryos expressed GFP gene showed mosaicism. They showed GFP expression at 1/4, 2/4 and 3/4 of blastomeres at the 4-cell stage. Among these 4-cell embryos, the expression rate of 1/4 blastomere group (54.6%) was higher than the other groups (15.3-30.7%). These results indicate that membrane disrupted sperm could attach with exogenous DNA, and that this procedure may be useful to introduce foreign gene into porcine oocytes. Therefore, our data suggest that the ICSI car be a useful tool to efficiently produce transgenic pig as well as other mammals.

• Korean Journal of Animal Reproduction
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• v.19 no.4
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• pp.271-281
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• 1996

1960년대에 본격적인 연구가 시도된 난세포질내 정자직접주입법(Intracytoplasmic Sperm Injection ; ICSI)은 1976년에는 Uehara 등에 의한 hamster의 연구에서 최초로 전핵의 형성에까지 성공하였다. 이후 계속된 연구를 통하여 여러 동물종에서 이 방법에 의한 난자의 수정 및 배발달에 성공하여, 1988년에 토끼, 1991년에 소, 1995년에는 생쥐에서 산자의 생산에 성공하였다. 한편, 사람에 있어서는 1988년 Lanzendorf 등에 의해 최초로 사람난자가 난세포질내 정자직접주입법에 의해 수정에 성공한 것이 보고되었으며, 1992년에는 Palermo 등에 의해 이 방법에 의해 수정된 수정란 이식을 통한 임신 및 성공적인 분만이 보고되었다. 난세포질내 정자직접주입법에 있어서는 정자의 운동성 및 첨체반응 등의 유무가 수정에 관여하는 중요한 요인이 아닌 것으로 알려지고 있으며, 성숙한 정자가 아닌 정소상체(미부-두부)정자 혹은 정소내의 미성숙정자를 사용하여 난세포질내 정자직접주입법을 시행하여도 수정 및 임신이 가능한 것으로 보고되었다. 또한 정자세포(spematid)나 원형정자세포(round spermatid)를 수정과 배발달이 관찰되었으며 사람에 있어서는 분만까지 성공하였다. 현재까지 이 난세포질내 정자직접주입법은 학술적으로는 난자-정자가 결합하는 기전을 밝히는 연구에 이용되어 왔으며, 임상적으로는 시험관아기시술에 있어서 정자의 기능, 수 등이 문제가 되어 수정이 어려운 남성불임환자에게 적용하여 좋은 성과를 거두어 왔다. 향후 이 기술은 유전자진단이나 남성불임환자의 처치에 폭넓게 사용될 것으로 기대된다.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.2
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• pp.130-134
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• 2020

Objective: The aim of this study was to compare the effects of conventional insemination (in vitro fertilization [IVF]) and intracytoplasmic sperm injection (ICSI) on the fertilization, developmental competence, implantation potential, and clinical pregnancy rate of embryos derived from in vitro matured oocytes of patients with polycystic ovary syndrome (PCOS). Methods: A prospective study was carried out among 38 PCOS patients who had undergone In vitro maturation (IVM) treatment. In total, 828 immature oocytes were collected from 42 cycles and randomly assigned for insemination by IVF (416 oocytes) or ICSI (412 oocytes). After fertilization, the embryos were cultured until the blastocyst stage and single embryos were transferred after endometrial preparation and under ultrasound guidance. Results: No significant differences were found in the maturation rate (78.1% vs. 72.6% for IVF and ICSI insemination, respectively; p= 0.076), fertilization rate (59.4% vs. 66.9% for IVF and ICSI insemination, respectively; p= 0.063), or the formation of good-quality blastocysts (40.9% vs. 46.5% for IVF and ICSI insemination, respectively; p= 0.314). Implantation and clinical pregnancy also did not show significant differences. Conclusion: There was a comparable yield of in vitro matured oocytes derived from PCOS patients in terms of fertilization, blastocyst formation, implantation rate, and clinical pregnancy between IVF and ICSI insemination. These findings provide valuable insights for choosing assisted reproductive treatment in women with PCOS, as IVM offers promising outcomes and is less invasive and less costly.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.1
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• pp.80-84
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• 2021

Objective: The aim of the study was to investigate pregnancy, obstetric, and neonatal outcomes in women with small (<4 cm) unilateral endometriomas. Methods: This retrospective study included 177 patients: 91 patients with small endometriomas and 86 controls with unexplained or tubal factor infertility who were treated at the Süleymaniye Gynecology and Maternity Training and Research Hospital Infertility Unit between January 2010 and July 2015. The groups were matched with regards to demographic characteristics such as age, body mass index, and infertility duration. All of the women in this study conceived via intracytoplasmic sperm injection. We compared pregnancy, obstetric, and neonatal outcomes between these groups. Results: Women with endometriomas had a higher biochemical pregnancy rate, but lower clinical pregnancy and live birth rates than women with unexplained and tubal factor infertility (p<0.05 for all). However no significant differences were found in terms of obstetric and neonatal complications between the two groups (p>0.05 for all). Conclusion: In this study, we found that women with endometriomas less than 4 cm were more prone to early pregnancy complications. We also showed that this group did not have any increased risks of late pregnancy, obstetric, and neonatal complications.