• Clinical and Experimental Reproductive Medicine
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• v.35 no.2
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• pp.131-141
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• 2008

Objective: To compare the outcomes of intracytoplasmic sperm injection (ICSI) with fresh and cryopreserved-thawed testicular spermatozoa in patients with azoospermia. Methods: One hundred and nine cycles (66 couples) where ICSI was planned with fresh or cryopreserved-thawed testicular spermatozoa were included in this study; Ninety two cycles (61 couples) with fresh testicular spermatozoa (fresh group) and seventeen cycles (13 couples) with cryopreserved-thawed testicular spermatozoa (cryopreserved-thawed group). We compared ICSI outcomes such as fertilization rate, implantation rate, pregnancy rate and miscarriage rate, which were statistically analyzed using Mann-Whitney U test or Fisher's exact test, where appropriate. Results: In 9 out of the 92 cycles where ICSI was planned with fresh testicular spermatozoa, testicular spermatozoa could not be retrieved. Fertilization rate tended to be higher in the fresh group than in the cryopreserved-thawed group ($58.0{\pm}27.8%$ vs. $45.9{\pm}25.0%$, p=0.076). The number of high quality embryos was significantly higher in the fresh group ($0.9{\pm}1.2$ vs. $0.2{\pm}0.5$, p=0.002). However, there were no significant differences in clinical pregnancy rate, implantation rate and miscarriage rate between the two groups. Conclusion: The results of this study suggest that although the use of cryopreserved-thawed testicular sperm for ICSI in patients with azoospermia may reduce fertilization capacity and embryo quality, it may not affect pregnancy rate, implantation rate and miscarriage rate. If testicular sperm can be obtained before ICSI procedure, the use of cryopreserved-thawed testicular sperm may also avoid unnecessary controlled ovarian hyperstimulation and cancellation of oocyte retrieval when spermatozoa cannot be retrieved as well as damage on testicular function by repeated TESE.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.3
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• pp.164-168
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• 2016

Objective: Assisted reproductive technology has been associated with an increase in multiple pregnancies. The most effective strategy for reducing multiple pregnancies is single embryo transfer. Beginning in October 2015, the National Supporting Program for Infertility in South Korea has limited the number of embryos that can be transferred per in vitro fertilization (IVF) cycle depending on the patient's age. However, little is known regarding the effect of age and number of transferred embryos on the clinical outcomes of Korean patients. Thus, this study was performed to evaluate the effect of the number of transferred blastocysts on clinical outcomes. Methods: This study was carried out in the Fertility Center of CHA Gangnam Medical Center from January 2013 to December 2014. The clinical outcomes of 514 women who underwent the transfer of one or two blastocysts on day 5 after IVF and of 721 women who underwent the transfer of one or two vitrified-warmed blastocysts were analyzed retrospectively. Results: For both fresh and vitrified-warmed cycles, the clinical pregnancy rate and live birth or ongoing pregnancy rate were not significantly different between patients who underwent elective single blastocyst transfer (eSBT) and patients who underwent double blastocyst transfer (DBT), regardless of age. However, the multiple pregnancy rate was significantly lower in the eSBT group than in the DBT group. Conclusion: The clinical outcomes of eSBT and DBT were equivalent, but eSBT had a lower risk of multiple pregnancy and is, therefore, the best option.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.139-144
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• 2001

This study was carried out to investigate the effects of the bull, sperm type and sperm pretreatment on the pronuclear formation and in vitro development after injection of spermatozoa into in vitro matured bovine oocytes. 1. Spermatozoa derived room four bulls(A, B, C and D) were used for ICSI. The male pronuclear formation and developmental rates were 73.9∼87.0% and 33.3∼60.9%, respectively. 2. The effects of sperm type were examined. Male pronuclear formation rates by using fresh-and frozen-sperm, tail-cutting and tail-scoring sperm were 82.0%. 78.0%. 42.2% and 51.1% (p<0.05) while development rates were 56.0%. 42.0%, 17.8% and 22.2%, respectively. Fresh sperm achived a high mail pronuclear- and development rates than those of other groups. 3. Cheroical pretreatments were tested and compared. When sperm were pretreated with heparin, BFF(bovine follicula fluid), His, Ca Ionophore(I) and I + caffeine, mate pronuclear formation and developmental rates were 66.7∼82.2% and 33.3∼60.6%. respectively. and these values of treatment of I + caffeine were higher than that of other methods. 4. The male pronuclear formation and developmental rates of oocytes obtained by ICSI treated with or without zona pellucida were 80.0%. 72.0% and 46.0%, 36.0%, respectively.

• Journal of Embryo Transfer
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• v.33 no.1
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• pp.23-30
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• 2018

The purpose of this study was to investigate the effects of the concentration of seminal plasma in aerobic and anaerobic conditions on the total motility(TM) and the progressive motility(PM) of spermatozoa in long term preservation of cooled equine semen. We also examine the pregnancy rates after artificial insemination using fresh, cooled or frozen semen, and different durations of cooled-preserved equine semen. In the aerobic state of cooled-preserved semen, As the increase of preserved duration to 24h, 48h, 72h, and 96h, TM tended to decrease in each of different concentrations of formalin-containing experimental group, TM tended to decrease regardless of the concentrations of SP. In different concentrations of SP, TM of without seminal plasma(SP W/O) group tended to be higher than that of SP 20%, SP 33% and SP 50%, especially TM of SP W/O group was significantly higher than other groups at 96 h (p<0.05). PM was higher in the groups of SP W/O and SP 20% than in the groups of SP 33% and SP 50% from 24 h to 72 h in cooled-preservation, especially PM of SP W/O group was significantly higher than other groups at 96 h (p<0.05). In the anaerobic condition of cooled-preserved semen, the results of TM and PM at different concentrations of SP were similar to the results in the aerobic condition although there was a difference in the ratio. The pregnancy rates of fresh-cooled, cooled-preserved and frozen semen were 66.3%, 60.7% and 34.5%, respectively, and the pregnancy rate of frozen semen was the lowest. We also found that it is possible to pregnancy after artificial insemination using 72 h cooled-preserved equine semen. There was similar of the pregnancy rates in the different month from April to August.

• Journal of Embryo Transfer
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• v.18 no.1
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• pp.27-33
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• 2003

The study was carried out to investigate the effects of morphology, reproductive cycle, incubation time and activation of oocytes in vitro maturation of canine oocytes and development of canine IVM/IVF embryos. The results were summarized as follows: 1. The developmental rates to 16 cells of fresh, salts and 4$^{\circ}C$-stored oocytes with and without cumulus cells were 14.3%, 5.0% and 7.5%, 2.8% and 5.7%, 0.0%, respectively. The rate of oocytes with cumulus cells(5.7%~14.3%) was higher than that of denuded oocytes(0.0%~5.0%). 2. The developmental rate to If cells of in vitro cultured oocytes recovered from ovaries collected at different stages of the reproductive cycle were 0.0%, 10.7%, 1.5%, respectively. 3. The developmental rate to 16 cells of fresh oocytes with cumulus cell cultured for 24, 32 and 48 hrs in $CO_2$ incubator were 0.0%, 5.3%, 11.8%, respectively. The rate of oocytes cultured for 48 hrs was higher than that oocytes cultured for 24 and 32 hrs. 4. The development to If cells treated activation and non-activation oocytes were 15.0%, 6.7%, respectively. The rate of oocytes treated activation was higher than that oocyte treat non-activation.

• Korean Journal of Plant Resources
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• v.32 no.1
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• pp.53-62
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• 2019

The purpose of this study was to establish the in vitro propagation system by shoot tip culture of six Hosta species native to Korea (Hosta capitata (Koidz.) Nakai, H. clausa Nakai, H. jonesii M.G.Chung, H. minor (Baker) Nakai, H. venusta F.Maek., and H. yingeri S.B.Jones) for mass proliferation and a new cultivar development. The shoot tips of each Hosta species were cultured on MS medium containing eight combinations of 0.5, 1.0, 2.0, 4.0 mg/L BA with 0.1 mg/L NAA, 0.1, 0.5, 1.0, 2.0 mg/L TDZ with 0.1 mg/L NAA, and without any PGRs (control). They were investigated on callus, somatic embryo, crown bud, differentiation and growth of shoot and root, total fresh weight after 8 weeks of culture. In all six Hosta species, callus and somatic embryo induction rate and multiple shooting rate of the PGRs treatment group were higher than that of the control group. The highest number of differentiated shoots were obtained on medium supplemented with 2.0 ㎎/L TDZ in H. capitata (5.4), 1.0 mg/L TDZ in H. clausa and H. jonesii (3.3 and 5.8, respectively), 0.5 mg/L BA in H. minor (11.1), 1.0 mg/L BA and 0.1 mg/L TDZ in H. venusta (8.1), and 0.5 mg/L TDZ in H. yingeri (9.8). In somatic embryo formation, the PGRs treatment group of H. jonesii and H. yingeri were more effective than the control group, and the effects were relatively less in H. capitata, H. clausa Nakai, H. minor, H. venusta. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. Crown bud formation of four Hosta species (H.capitata, H. clausa, H. jonesiig, and H. yingeri) were also higher in the PGRs treatment group than in the control group. H. clausa showed no significant effect on callus and shoot differentiation regardless of the type and concentration of cytokinin, but slightly increased in formation of crown bud in TDZ.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.1
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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.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.4
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• pp.178-188
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• 2019

Objective: To determine the clinical pregnancy (CP) and live birth (LB) rates arising from frozen embryo transfers (FETs) that had been generated under the influence of in vitro fertilization (IVF) adjuvants given to women categorized as poor-prognosis. Methods: A registered, single-center, retrospective study. A total of 1,119 patients with first FETs cycle include 310 patients with poor prognosis (109 treated with growth hormone [GH], (+)GH group vs. 201 treated with dehydroepiandrosterone, (-)GH group) and 809 patients with good prognosis (as control, (-)Adj (Good) group). Results: The poor-prognosis women were significantly older, with a lower ovarian reserve than the (-)Adj (Good) group, and demonstrated lower chances of CP (p< 0.005) and LB (p< 0.005). After adjusting for confounders, the chances of both CP and LB in the (+)GH group were not significantly different from those in the (-)Adj (Good) group, indicating that the poor-prognosis patients given GH had similar outcomes to those with a good prognosis. Furthermore, the likelihood of LB was significantly higher for poor-prognosis women given GH than for those who did not receive GH (p< 0.028). This was further confirmed in age-matched analyses. Conclusion: The embryos cryopreserved from fresh IVF cycles in which adjuvant GH had been administered to women classified as poor-prognosis showed a significant 2.7-fold higher LB rate in subsequent FET cycles than a matched poor-prognosis group. The women with a poor prognosis who were treated with GH had LB outcomes equivalent to those with a good prognosis. We therefore postulate that GH improves some aspect of oocyte quality that confers improved competency for implantation.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.1
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• pp.47-51
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• 2017

Objective: This study was designed to report the status of assisted reproductive technology (ART) therapy in South Korea between January 1, 2012 and December 31, 2012. Methods: A localized online survey, originally developed by the International Committee Monitoring Assisted Reproductive Technologies, was first launched and provided to all available ART centers via email in 2015. Fresh embryo transfer (FET) cases were categorized as standard in vitro fertilization, intracytoplasmic sperm injection (ICSI), or half-ICSI. Thawed embryo transfer (TET) and other related procedures, including surgical sperm retrieval, were surveyed. Results: Data from 33,956 ovum pick-up procedures were provided by 75 clinics in 2012. Of the 33,088 cycles in which ovums were retrieved, a complete transfer was performed in 90.5% (29,932 cycles). In addition, 10,079 FET cycles were confirmed to have resulted in clinical pregnancy, representing a pregnancy rate of 30.5% per ovum pick-up and 33.7% per ET. The most common number of embryos transferred in FET was 2 (41.6%), followed by 3 (34.0%), and non-elective single ETs (10.0%). Of the 10,404 TET cycles in which transfer was completed, 3,760 clinical pregnancies (36.1%) were confirmed by ultrasonography. Conclusion: The overall clinical pregnancy rate for FET and TET cycles in 2012 was higher than in 2011 (33.7% vs. 33.2% and 36.1% vs. 31.1%, respectively). The most common number of embryos transferred in FET cycles was 2, unlike in 2011.

• Journal of Embryo Transfer
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• v.27 no.3
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• pp.183-187
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• 2012

Adult stem cell transplantation has been increased every year, because of the lack of organ donors for regenerative medicine. Therefore, development of reliable and safety cryopreservation and bio-baking method for stem cell therapy is urgently needed. The present study investigated safety of dimethyl sulfoxide (DMSO) such as common cryoprotectant on porcine bone marrow derived mesenchymal stem cells (pBM-MSCs) by evaluating the activation of Caspase-3 and -7, apoptosis related important signal pathway. pBM-MSCs used for the present study were isolated density gradient method by Ficoll-Paque Plus and cultured in A-DMEM supplemented 10% FBS at $38.5^{\circ}C$ in 5% $CO_2$ incubator. pBM-MSCs were cryopreserved in A-DMEM supplemented either with 5%, 10% or 20% DMSO by cooling rate at $-1^{\circ}C$/min in a Kryo 360 (planner 300, Middlesex, UK) and kept into $LN_2$. Survival rate of cells after thawing did not differ between 5% and 10% DMSO but was lowest in 20% DMSO by 0.4% trypan blue exclusion. Activation of Caspase-3 and -7 by Vybrant FAM Caspase-3 and -7 Assay Assay Kit (Molecular probes, Inc.OR, USA) was analyzed with a flow cytometer. Both of cryopreserved and control groups (fresh pBM-MSCs) were observed after the activation of Caspase-3 and -7. The activation did not differ between 5% and 10% DMSO, but was observed highest in 20% DMSO. Therefore 5% DMSO can be possibly used for cell cryopreservation instead of 10% DMSO.