• Clinical and Experimental Reproductive Medicine
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• v.47 no.4
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• pp.293-299
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• 2020

Objective: The goal of the present study was to investigate the rate of chromosomal aneuploidies in surplus embryos after sex determination at the cleavage stage. Then, the same chromosomal aneuploidies were evaluated in blastocysts after extended culture. Methods: Sixty-eight surplus embryos were biopsied at the cleavage stage and incubated for an additional 3 days to allow them to reach the blastocyst stage. The embryos were reanalyzed via fluorescence in situ hybridization (FISH) to examine eight chromosomes (13, 15, 16, 18, 21, 22, X, and Y) in both cleavage-stage embryos and blastocysts. Results: Although the total abnormality rate was lower in blastocysts (32.35%) than in cleavage-stage embryos (45.58%), the difference was not significant (p=0.113). However, when we restricted the analysis to autosomal abnormalities, we observed a significant difference in the abnormality rate between the cleavage-stage embryos (44.11%) and the blastocysts (17.64%, p=0.008). A higher rate of sex chromosomal abnormalities was also observed in cleavage-stage embryos (29.4%) than in blastocysts (14.70%, p=0.038). Conclusion: The data indicated that embryo biopsy should be conducted at the blastocyst stage rather than the cleavage stage. The results also emphasized that examination of common chromosomal aneuploidies apart from sex selection cycles can be conducted in the blastocyst stage with the FISH method.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.2
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• pp.103-108
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• 2011

Objective: To investigate adverse pregnancy outcomes in non-obese women with polycystic ovary syndrome (PCOS) compared with obese-PCOS and control groups. Methods: Women with PCOS who underwent assisted reproductive technology (ART) from August, 2003 to December, 2007, were considered. A total of 336 women with PCOS were included in the study group and 1,003 infertile women who had tubal factor as an indication for ART were collected as controls. They were divided into four groups: a non-obese PCOS group, obese-PCOS group, non-obese tubal factor group, and obese tubal factor group, with obesity defined by a body mass index over 25 kg/$m^2$, and reviewed focusing on the basal characteristics, ART outcomes, and adverse pregnancy outcomes. Results: There was no difference among the groups' the clinical pregnancy rate or live birth rate. Regarding adverse pregnancy outcomes, the miscarriage rate, multiple pregnancy rate, and prevalence of preterm delivery and pregnancy induced hypertension were not different among the four groups. The incidence of small for gestational age infant was higher in the PCOS groups than the tubal factor groups ($p$ <0.02). On the other hand, the morbidity of gestational diabetes mellitus (GDM) was not high in the non-obese PCOS group but was in the obese groups. And in the obese PCOS group, the newborns were heavier than in the other groups ($p$ <0.02). Conclusion: Non-obese PCOS presents many differences compared with obese PCOS, not only in the IVF-parameters but also in the morbidity of adverse pregnancy outcomes, especially in GDM and fetal macrosomia.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.2
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• pp.121-129
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• 2001

Objective: The aim of this study is to compare the efficiency of a method for the cryopreservation of mouse blastocyst.. Methods: Mouse embryos were obtained at 2-cell stage and cultured to blastocyst stage in T6 medium supplemented with 10% fetal bovine serum. Morphologically normal blastocysts were collected and randomly divided to one control and four experimental groups. In control group, blastocysts were cultured in vitro continuously for additional two days. In group 2, blastocysts were exposed to vitrification solution (ethylene glycol) only without cryopreservation (exposure only group). In group 3, 4 and 5, blastocysts were cryopreserved by slow-freezing procedure with glycerol (slow-fteezing group) or by vitrification procedure using EM grids (EM grids group) and cryoloop (cryoloop group), respectively. Frozen blastocysts were thawed and cultured for additional two days. Twenty four hours after thawing, some blastocysts were fixed and stained with Hoechst 33342 (bisbenzimide) and the number of nuclei in each blastocysts were counted to confirm the survival of bias to cysts in experimental groups. Results: Survival rate and hatching rate of the blastocysts in slow-freezing group (24 h: 72.4% and 66.0%, 48 h: 63.2% and 64.6%) and EM grids group (24 h: survival rate 77.3%, 48 h: 70.1% and 71.4%) were significantly lower ($X^2$-test p<0.05) than those of control group (24 h: 93.4% and 86.0%, 48 h: 88.5% and 90.7%). In contrast, the survival rate and hatching rate of the blastocysts in cryoloop group (24 h: 84.1% and 84.1%,48 h 79.3% and 87.7%) is well compared with those in the control group. The mean (${\pm}SD$) cell number of blastocyst in the exposure only ($89.2{\pm}11.5$), EM grids ($85.0{\pm}10.3$) and cryoloop ($89.0{\pm}11.0$) groups, except slow-freezing group ($79.0{\pm}10.0$), were not significantly different from that of control group ($93.1{\pm}13.9$) 24 h after thawing (Student's t-test). Conclusion: This study demonstrates that higher survival rate of vitrified-thawed mouse blastocyst can be obtained using cryoloop as the embryo container at freezing rather than slow-freezing or vitrification using EM grids. The results of this study suggest that vitrification using cryoloop (with ethylene glycol) may be a preferable procedure for mouse blastocyst cryopreservation and could be applied to the human blastocyst cryopreservation.

• Journal of Veterinary Clinics
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• v.21 no.2
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• pp.115-121
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• 2004

The purpose of this study was to ascertain the breeding efficiency in Thoroughbred mare. A total of 106 mares were investigated for the status of follicle (462 cases), ovulation (179 cases) and pregnancy (346 cases). Of total examination, 46.8% was follicle measure to determine breeding time, and mating rate per cases examined was 39.9%. There was no correlation between reproductive results and size of follicles or endometrial edema or degrees of teasing alone. 143 cases were ovulated among 179 cases which were performed ovulation examination, and ovulation rate and fertilization rate per mating times were 79.9% and 39.0%, respectively. The use of hCG(human chorionic gonadotropin), to facilitate ovulation, presented to increase occurrence of double ovulations and twin fertilizations In conclusion, though more examination to estimate the optimal breeding time and higher mating rate was performed, fertilization rate per mating times was lower and then reproductive efficiency also became decreased. Therefore, it seemed that accurate examination of reproductive tracks, appropriate teasing programme and hCG administration before ovulation were of help to improve ovulation rate and fertilization rate.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.261-267
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• 1992

The reproductive performance of SPF ICR mice under single paired mating were examined to get reproductive background data and to establish single paired rotational mating system. The results obtained were as follows : average litter size was 15.4$\pm$2.0 heads ; average weaning rate was 95.7$\pm$4.9% ; sex ratio(male/female) was 1.09$\pm$0.26 ; aveage delivery interval was 23.0$\pm$2.4 days. It was given the largest litter size at the age of 121~150 days and in 2nd~4th parities, but at the age of under 90 days and in 1st parity weaning rate and delivery interval were higher and shorter than those of the other ages and parities, respectively. In sex ratio, the number of male litters was slightly increased from that of female litters. The weaning rate of litters from dams which nuresed 12 litters was the highest among those of different litter sizes, and it was decreased dependent upon increment of litter size. There were no difference among 4 groups for reproductive performance, therefore the present study could have important sources for animal breeders who produce mice using the single paired rotational mating system.

• Clinical and Experimental Reproductive Medicine
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• v.44 no.1
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• pp.52-55
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• 2017

The aim of this study was to report a successful pregnancy using completely immotile frozen-thawed spermatozoa selected by laser. A single laser shot was used to detect the presence of viable immotile spermatozoa in fresh and frozen-thawed testicular spermatozoa. The viability rate was 55.8% after the laser detection, and cryopreservation was carried out immediately. The thawing test was performed on the day of oocyte pick-up, and no motile sperm were observed after extending the culture for another 4 hours, while a survival rate of 39.8% was detected using the laser. In all, five mature oocytes were injected, resulting in four cases of normal fertilization (80%) on day 1. Further, two high-quality day 3 embryos were transferred, which resulted in a singleton pregnancy. Our study demonstrates that completely immotile spermatozoa are worth cryopreserving for further intracytoplasmic sperm injection, which provides a new insight into male fertility preservation in cases of completely immotile spermatozoa.

• Journal of Animal Environmental Science
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• v.1 no.1
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• pp.65-75
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• 1995

This experiment was carried out to the effects of population density and C/N ratios of feed on the growth, reproductive effciency and cast producation of the earthworm(Eisenia foetida). The population densities of 50, 100, 150, 200 and 250 individuals of the earthworm fed with different C/N ratios of 25, 35, 45 and 55 cow manures were studied in rearing box($6,400cm^3$), and at the fertility stage during a period of 60 days. The results were summarized as follows; The survial rate(SR), increasing rate(IR), reproductive efficiency(RE) and cast production of the earthworms showed highest values in C/N ratio of 25. These results may indicate that C/N ratio of 25 is a very favourable feed for the growth of the earthworms. The survial rate(SR) indicated significant positive correlation with reproductive efficiency(RE) in different C/N ratios of feeds. The survial rate(SR) showed highest values in population densities of $50{\sim}100$ worms/$6,400cm^3(64.0{\sim}128.0cm^3/worm$). On the contrary, increasing rate(IR) tended to decreased with the increased population densities. The survival rate(SR) indicated significant negative correlation with reproductive efficiency(RE) in different population densities of the earthworms. The cast production estimated were $31.6mg{\sim}67.4mg/day/worm$ grown in optimum population densities($50{\sim}100\;worms/6,400cm^3$). The earthworm casting are an excellent soil conditioning material or organic fertilizer sources with a high chemical composition and their physical properties.

• Clinical and Experimental Reproductive Medicine
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• v.45 no.3
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• pp.110-115
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• 2018

Objective: To compare our in-house method of embryo freezing with Cryotop vitrification in terms of immediate survival, subsequent cleavage and blastocyst formation, and cell numbers in blastocysts. Methods: Two-cell mouse embryos were randomly allocated into three groups: a non-frozen control group (group 1, n = 300), a group that underwent Cryotop vitrification (group 2, n = 300), and a group that underwent our in-house freezing method (group 3, n = 300). Results: There were no significant differences between groups 2 and 3 in the immediate survival rate (96.3% vs. 98.6%, respectively; p= 0.085), the further cleavage rate (91.7% vs. 95.0%, respectively; p= 0.099), or the blastocyst formation rate (80.7% vs. 78.6%, respectively; p= 0.437). The cell numbers in the blastocysts from groups 1, 2, and 3 were comparable ($88.99{\pm}10.44$, $88.29{\pm}14.79$, and $86.42{\pm}15.23$, respectively; p= 0.228). However, the percentage of good-quality blastocysts in the Cryotop vitrification group was significantly higher than in the group in which our in-house method was performed, but was lower than in the control group (58.0%, 37.0%, and 82.7%, respectively; p< 0.001). Conclusion: At present, our method is inferior to the commercial Cryotop vitrification system. However, with further improvements, it has the potential to be useful in routine practice, as it is easier to perform than the current vitrification system.

• 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.45 no.3
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• pp.129-134
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• 2018

Objective: In frozen and thawed embryos, the zona pellucida (ZP) can be damaged due to hardening. Laser-assisted hatching (LAH) of embryos can increase the pregnancy rate. This study compared thinning and drilling of the ZP before frozen embryo transfer (FET). Methods: Patients were randomly allocated into two groups for LAH using thinning or drilling on day 2 after thawing. Twenty-five percent of the ZP circumference and 50% of the ZP thickness was removed in the thinning group, and a hole $40{\mu}m$ in diameter was made in the drilling group. Results: A total of 171 in vitro fertilization/intracytoplasmic sperm injection FET cycles, including 85 cycles with drilling LAH and 86 cycles with thinning LAH, were carried out. The thinning group had a similar ${\beta}$-human chorionic gonadotropin-positive rate (38.4% vs. 29.4%), implantation rate (16.5% vs. 14.4%), clinical pregnancy rate (36.0% vs. 25.9%), miscarriage rate (5.8% vs. 2.4%), ongoing pregnancy rate (30.2% vs. 23.5%), and multiple pregnancy rate (7.0% vs. 10.6%) to the drilling LAH group. There were no significant differences in pregnancy outcomes between subgroups defined based on age (older or younger than 35 years) or ZP thickness (greater or less than $17{\mu}m$) according to the LAH method. Conclusion: The present study demonstrated that partial ZP thinning or drilling resulted in similar outcomes in implantation and pregnancy rates using thawed embryos, irrespective of women's age or ZP thickness.