• 한국수정란이식학회지
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• 제18권2호
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• pp.157-161
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• 2003

본 연구는 소형 고양이의 불임 해결과 체외수정란을 생산하기 위한 방안으로서 난자의 형태, 번식주기, 배양시간 및 활성화 처리가 난포란의 체외수정 및 체외발생에 미치는 영향을 조사하였다. 1. 신선 및 salt에 보존한 난소로부터 회수한 난구세포부착 및 나화 난자를 각각 배양했을때 체외수정율 및 분할율은 65.7%와 17.1%, 28.6%와 8.6% 및 57.1%와 13.3%, 23.3%와 3.3%로서 난구세포 부착 신선난자가 나화 난자에 비해 높은 체외발생률을 나타냈다. 2. 휴지기, 발정기 및 황체기 단계로 구분하여 채취한 난포란을 성숙배양 후 수정시켰을 때 체외수정율은 각각 68.9%, 44.4%, 48.9%였으며, 분할율은 각각 17.8%, 8.9%, 12.8%로 나타났다 3. 24, 36 및 48시간 각각 배양한 난포란을 성숙 배양 후 수정시켰을 때 체외수정율은 각각 66.7%, 46.7%, 48.9%였으며, 분할율은 각각 17.8%, 11.1%, 8.5%로 나타났다. 4. 난자를 활성화처리 및 비활성화처리 후 각각 체외수정시켰을 때 체외수정율은 각각 57.4%와 31.4%였고, 체외분할율은 22.9%와 11.4%로서 활성화 처리를 한 난자가 높은 체외발생율을 나타냈다.

• 한국수정란이식학회지
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• 제31권3호
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• pp.185-190
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• 2016

During the ovary preservation in low temperature, the cumulus oocyte complexes(COCs) lose their developmental competences after in vitro fertilization. We used phosphate-buffered saline (PBS) as a basic solutions of at various temperatures (25, 15 or $5^{\circ}C$) and supplemented them with 1mM glucose and 0.5mM glutamine as a source of carbohydrate metabolites. After recovery of COCs and in vitro fertilization, a significantly higher number of oocytes developed into blastocysts. The developmental competence of embryos that were originated from ovaries preserved at $15^{\circ}C$ was increased compared to those of 25 or $5^{\circ}C$. The maturation rate of oocytes was not differed between 24 and 36 h at $15^{\circ}C$ but showed lower than control group (71% versus 78%). In vitro-fertilized oocytes from ovaries stored at $25^{\circ}C$ for 24 h or at $5^{\circ}C$ for 24 h had a significantly decreased developmental potentials, but at $15^{\circ}C$ did not (27% versus 29% of blastocysts to develop into day 8). With these results, bovine ovaries can be preserved at $15^{\circ}C$ for 36 h without decreasing developmental capacity of in vitro-fertilized oocyte at least to the blastocyst stage. This information provides valuable information of preserving ovaries for embryo transfer or in vitro embryo production.

• Clinical and Experimental Reproductive Medicine
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• 제45권4호
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• pp.189-194
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• 2018

Objective: The aim of this study was to compare the rate of maturation, fertilization, and embryo development of in vitro-matured human oocytes derived from pregnant and non-pregnant women. Methods: Immature oocytes were obtained by needle aspiration from 49 pregnant women (group 1) who underwent a cesarean section at term and 77 non-pregnant women (group 2) who underwent a gynecological operation during the same period (8 months). Healthy immature oocytes (530 in group 1 and 539 in group 2) were cultured and assessed for maturation 36 hours later. Mature oocytes were inseminated by intracytoplasmic sperm injection and cultured up to 144 hours. Results: The percentage of degenerated oocytes was significantly higher (12.1% vs. 6.3%; p<0.001) in group 1 than in group 2. There was no significant difference in the maturation rate (66.8% vs. 68.1%; p=0.698), fertilization rate (66.7% vs. 67.6%; p=0.857), or the rate of formation of good-quality blastocysts (46.2% vs. 47.2%; p=0.898) in oocytes obtained from pregnant and non-pregnant women. Conclusion: The developmental competence of immature oocytes did not differ between pregnant and non-pregnant women.

• 한국수정란이식학회지
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• 제12권1호
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• pp.1-10
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• 1997

The present study was conducted to investigate the influence of embryo cell stage at 18h post-fusion and oocyte preactivation on sebsequent in vitro developmental potential in the nuclear transplant rabbit embryos. The embryos of 16-cell stage were collected and synchronized to G$_1$ phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome rnass from the oocytes collected by non-dis-ruptive microsurgery procedure. The separated G$_1$ phase blastomeres of 32-cell stage were injected into non-preactivated recipient cytoplasms. Otherwise, the enucleated recipient cytoplasms were preactivated by electrical stimulation at 18h post-hCG injection and the separated G$_1$ phase blastomeres of 32-cell stage were injected. Mter culture until 20h post-hOG injection, the nuclear transplant oocytes were electrofused by electrical stimulation. The fused nuclear transplant embryos were classified into 3~4-cell, 2-cell and 1-cell stage at 18 hrs post-fusion and cultured until the embryos reached blastocyst stage. The developmental rate to blastocyst stage was significantly (P <0.05) higher in all the reconstituted embryos of 3~4-cell stage(58.0%) than in 2 and icell stage. The developmental rate to blastocyst stage in the embryos of 3~4-cell stage at 18 hrs post-fusion was significantly (P<0.05) higher in the reconstituted without oocyte preactivation(77.8%) than in the oocyte-preactivated embryos (33.3%). These results indicated that the higher rate of in the in vitro development to blastocyst stage might be obtained form the embryos which were reconstituted with nuclear donor of G$_1$ phase and non-preactivated oocyte, and developed more rapidly for 18 hrs post-fusion.

• 한국가축번식학회지
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• 제21권3호
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• pp.219-228
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• 1997

To examine the efficiency of nuclear transplantation the influence of electrical preactivation of recipient cytoplasm on the in vitro developmental potentyl in the nuclear transplant rabbit embryos were evaluated. The embryos of 16-cell stage were collected and synchronized to G1 phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome mass by non-disruptive microsurgery procedure. The separated G1 phase blastomeres of 32-cell stage were put into the non-preactivated and/or the preactivated recipient cytoplasm by electrical stimulation. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused. The fused nuclear transplant embryos were co-cultured with rabbit oviduct epithelial cells and monitored every 24h to assess for developmental rate. After in vitro culture for 120h, the nuclear transplant embryos developed to blastocyst stage were stained with Hoechst 33342 and their blastomere were counted. The electrofusion rate was similar to the non-preactivated and preactivated recipient cytoplasm(81.8 and 85.7%, respectively). However, the in vitro developmental rate to blastocyst stage with the non-preactivated recipient cytoplasm (57.1%) was found significantly (P<0.05) higher, compared to the preactivated recipient cytoplasm(20.8%). The cell counts of nuclear transplant embryos developed to blastosyst stage were increased significantly (P<0.05) more in the non-preactivated recipient cytoplasm (163.7 cells), as compared with the preactivated recipient cytoplasm(85.4 cells). These results considered better that non-preactivated oocytes, MII phase oocytes, were used for recipient cytoplasms in the rabbit nuclear transplant procedure.

• 한국수정란이식학회지
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• 제9권1호
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• pp.73-83
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• 1994

The objective of this study was to develop an effective in vitro production system capable of obtaining more porcine embryos from immature oocytes. These experiments were thus conducted to examine the effect of oocytes type and maturation time on the in vitro maturation(IVM) and fertilization(IVF) of oocytes and the in vitro development (IVD)of IVF embryos. 1. The degree of oocyte maturation based on cumulus expansion index(GEI) did not differ for A- and B-typed oocytes but the index of oocyte type C was lower(P<0.05) than that of other oocyte types. 2. When the oocytes of type A and B were matured for 36, 42 and 48hrs, the GEl was not different between the 36- and 42-h maturation but the GEl after 48hrs was greatly lower(P<0.05) than that of other maturation times. 3. The highest cleavage rate(48.6%) of IVF oocytes was obtained from A typed oocytes and 42-h maturation but the developmental potential based on cleavage index was the highest when B-typed oocytes were matured for 42hrs.

• 한국수정란이식학회지
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• 제9권1호
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• pp.85-93
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• 1994

This study was carried out to investigate the effects of gonadotropins added during maturation of porcine oocytes on the in vitro maturation(IVM), in vitro fertilization(IVF) and developmental potential of embryos. The follicular oocytes were cultured in TCM-199 medium containing different combination of gonadotropins(5$\mu$g /ml FSR or 1OIU /ml PMSG and 1O$\mu$g /ml LH or 1OIU /ml hCG), 10% FCS and 10% PFF for 36~48h in a incubator with 5% $CO_2$ in Air at 39$^{\circ}C$ and then matured oocytes were again cultured to 120h after IVF for 6~7h with heparin(100$\mu$g /m')-treated sperm. When the oocytes were matured for 42brs in the medium containing FSH+LH, FSH+hCG, PMSG+LH or PMSG+hCG, the JVF rate of each treatment was 50.0%, 52.9%, 66.7% and 70.0%, respectively. The highest CEI (cumulus cell expansion index) was obtained from PMSG+hCG-added medium and the highest polyspermic penetration resulted from FSH+LH-added medium. The cleavage of IVF oocytes derived from hormone added IVM was significantly(P<0.05) promoted by PMSG+hCG and the cleavage rate after 36-h, 42-h and 48-h maturation aws 53.0%, 56.7% and 45.6%, respectively. The highest developmental potential resulted from the oocytes derived from PMSG+LH -added IVM.

• Clinical and Experimental Reproductive Medicine
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• 제47권2호
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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.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.68-68
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• 2001

Blastocyst formation, consisting of the inner cell mass (ICM) and trophectoderm (TE), is the first differentiation process during embryonic development in mammals. It has been hypothesized that the proportion of ICM to TE in the blastocyst may be crucial for subsequent developmental competence of early embryos, which it may be expressed as a sensitive indicator for evaluating in vitro systems. In this study ICM/total cell ratio of nuclear transfer (NT) embryos was compared with IVF-derived and in vivo embryos. Somatic cell nuclei obtained from a fetus at Day 40 of gestation were transferred into the enucleated oocyte and then cultured in NCSU 23 medium for 6 days as previously described (Koo et al., Biol. Reprod. 2000; 63:986-992). ICM and TE cells of blastocysts were determined by using a differential staining method (Han et al., Biol. Reprod. 1999; 60:1110-1113). Development rate (9.8$\pm$2.5%, 23/225) to the blastocyst stage of NT embryos was lower than IVF embryos (23.8$\pm$2.7%, 53/223). Thus, a difference was detected in the in vitro developmental rate to blastocyst stage between NT and IVF-derived embryos (P<0.05). In the next experiment, we investigated ICM and TE nuclei to assess the quality of blastocysts that produced by NT, IVF and in vivo, respectively. NT blastocysts (27.6$\pm$8.3) showed a smaller total cell number than IVF-derived (42.6$\pm$17.4) and in vivo embryos (283.9$\pm$103.5) (P<0.05). Ratios of ICM/total cells in NT, IVF and in vivo blastocysts were 15.1$\pm$ 18.6% (n=56), 12.3$\pm$9.2% (n=57) and 30.4$\pm$6.8% (n=40), respectively. Individual blastocysts for the ratio of ICM/total cells were assigned to 3 groups (I; <20%, II; 20 to 40% and III;>40%). As the results, most in vivo blastocysts (97.5%, 39/40) were distributed into group II while most NT (78.6%, 44/56) and IVF-derived blastocysts (82.5%, 47/57) were allocated to group I. Thus, our data show that NT or IVF-derived embryos have aberrant morphology during early development in vitro systems, suggesting that these anomalies may result in developmental failures of the NT embryos to term.

• 한국수정란이식학회지
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• 제18권1호
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• pp.27-33
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• 2003

본 연구는 소형 개의 불임 해결과 체외수정란을 생산하기 위한 방안의 하나로써 난자의 형태, 번식주기, 배양시간 및 활성화 처리가 난포란의 체외성숙 및 체외발생에 미치는 영향을 조사하였다. 1. 신선, salt 및 4$^{\circ}C$에 보존한 난소로부터 채취한 난구세포부착 난자와 나화 난자로 각각 체외수정시켰을 때 16세포기로의 발생율은 14.3%, 5.0% 및 7.5%, 2.8%, 5.7% 및 0.0%로써 난구세포 부착난자군의 체외발생율이 나화 난자군에 비해 높게 나타났다. 2. 발정주기를 inactive, follicular, luteal 단계로 구분하여 채취한 난포란을 각각 체외배양시켰을 때 GV 및 MII로의 발생율은 11.3%와 9.4%, 50.7%와 26.7%, 16.9%와 13.8%였고, 16세포기로의 체외발생율은 0.0%, 10.7%, 1.5%였다. 3 신선한 난구세포 부착 난자를 각각 24, 32, 48시간 성숙배양 후 체외수정시켰을 때 분할율은 8.6%, 15.8%, 23.5%였으며, 16세포기로의 체외발생율은 각각 0.0%, 5.3%, 11.8%로써 48시간 배양군이 가장 높은 발생율을 나타냈다. 4. 활성화 처리 및 비활성화 처리 난자를 각각 체외수정시켰을 때 분할율은 각각 42.5%, 22.2%였고 16세포기로의 체외발생율은 각각 15.0%, 6.7%로써 활성화 처리 난자군이 비활성처리 난자군에 비해 높은 발생율을 나타냈다.