• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2003년도 추계학술대회
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• pp.157-157
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• 2003

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2005년도 제5회 발생공학 국제심포지엄 및 학술대회
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• pp.44-45
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• 2005

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2003년도 제3회 발생공학 국제심포지움 및 학술대회
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• pp.66-66
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• 2003

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2006년도 The 6th Intermational Symposium on Developmental Biotechnology
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• pp.88-88
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• 2006

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2004년도 제4회 발생공학 국제심포지움 및 학술대회
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• pp.69-69
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• 2004

• 한국수정란이식학회지
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• 제10권3호
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• pp.209-217
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• 1995

The present study was carried out to develop a cloning technology of mouse embryos by nuclear transplantation with electrofusion and to produce cloned offsprings by transfer of reconstituted embryos. A single nucleus from two- and eight-cell embryos was transplanted into the enucleated two-cell embryos by rnicromanipulation. The fusion of nucleus with recipient cytoplasm and the subsequent development of reconstituted embryos in vitro as well as in vivo to term were examined to determine the optimal electrofusion parameters for nuclear transplantation in mouse embryos. The successful enucleation of donor embryos was 84.9 and 83.3% in two- and eight-cell stage, respectively, and the successful injection of nucleus from two- and eight-cell donor embryos into the perivitelline space of enucleated two-cell embryos were 85.1 and 84.7%, respectively. No significant differences were found in enucleation or injection rate between the cell stages of donor embryos. When the blastomeres of intact two-cell mouse embryos were electrofused in 0.3 M mannitol medium(100 $\mu$sec., 3 pulses), the fusion rate was similarly 93.2, 92.2 and 92.0% in 1.0, 1.5 and 2.0 kV /crn, respectively, but in vitro development to blastocyst of the fused two-cell embryos was significantly(P<0.05) lower in 2.0 kV/cm (63.4%) than in 1.0 kV/cm (91.7%) or 1.5 kV/cm (82.4%). The development in vitro to eight-cell stage of the reconstituted embryos with nucleus from two-cell stage(45.5%) was significantly(P<0.05) higher than that from eight-cell stage blastomeres (16.7%). The number of blastomeres of the intact embryos at blastocyst stage was 50i0.6 and 55$\pm$2.4 in in vitro and in vivo cultured mouse embryos, respectively, but significantly(P<0.05) decreased to 35$\pm$0.7 in nuclear transplanted blastocyst embryos. The conception rate of mice following embryo transfer was 32.1% in the reconstituted two-cell embryos using two-cell donor nuclei, which was comparable to the fresh two-cell embryos(40.6%). However, the rate of development in vivo to term following embryo transfer of the reconstituted two-cell embryos using two-cell donor nuclei (23.5%) was significantly(P<0.05) lower compared with the percentage of two-cell fresh embryos(31.5%).

• 한국동물학회지
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• 제39권4호
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• pp.352-361
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• 1996

내인성 암원유전자인 c-myc유전자는 세포의 증식과 분화에 밀접한 연관되어 있으며, 그의 생물학적인 기능은 이형결합체인 myn과의 결합을 통해서 이루어진다. 본 연구에서는 착상전 생쥐 배아에서의 내인성 c-myc 유전자와 myn 유전자의 발현을 조사하기 위하여 RT-PCR 방법을 이용하였다. myn 유전자 산물은 배아 발생시기를 거쳐서 균일하게 발현된 반면,c-myc 유전자의 발현은 차후 포배기 시기까지 상당한 양으로 증가1세포기에 측정된 후,2세포기에 들어 현저하게 줄었으나, 차후 포배기 시기까지 상당한 양으로 증가하였다. 이러한 c-myc과 myn유전자발현의 비균등한 조화가 중요한 역할을 할 것으로 사료된다. 착상전 생쥐 초기배아 발생 과정에서의 myn 유전자의 기능을 알아 보고자, myn에 대한 antisense oligouncleotides(Myn2, Myn3)를 1세포기이 수정란에 미세주입하였다. Myn2와 Myn3의 미세주입에 의해 상실기/포배기의 변이 시기에 비정상적인 발생이 야기되었다. 이사의 결과로, c-myc은 그의 이형결합체인 myn과 함께 착상전 생쥐 초기배아에서의 중요한 역할을 할 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제24권2호
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• pp.233-239
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• 1997

Embryos of most mammalian species grown in vitro would undergo developmental arrest at the approximate time of genomic activation. Stage-specific cell block and the resulting rapid loss of embryo viability in conventional culture media have limited the duration for which embryos may be cultured prior to transfer. As a result, embryos are usually transferred to the uterus at the 4-to 8-cell stage to avoid the loss of viability associated with long-term in vitro culture. Early transfer has led to asynchrony of the endometrium-trophectoderm interaction at the time of implantation and a resultant reduction in the rate of implantation. To overcome these problems, a variety of co-culture systems has been devised in which embryos can develop for a longer period prior to embryo transfer. Vero cells, derived from African green monkey kidney, share a common embryologic origin with cells from the genital tract. In addition, they are potentially safe to use, since they are highly controlled for viruses and other contaminants. Therefore, co-culture using Vero cells has been widely utilized to enhance embryo viability and development, although not without controversies. We thus designed a series of experiments to demonstrate whether Vero cells do indeed enhance mouse embryo development as well as to compare the efficacy of co-culturing mouse 1-cell embryos on Vero cell monolayer in both Ham's F-10 and human tubal fluid (HTF) culture media. 1-cell stage ICR mouse embryos were cultured either in the presence of Vero cells (Group A) or in conventional culture medium alone (Group B). In Ham's F-10 significantly more 3-to-8cell embryos developed in group A than group B (59.8 versus 10.0%; p<0.01). In contrast, there was no significant difference in embryonic development both group A and group B in HTF. However, significant differences were noted only in later embryonic stage (13 and 0%; p<0.05 of group A and B respectively, hatching or hatched). In Ham's F-10, we also could observe the beneficial effect of Vero cell on hatching process (70.7 and 42.1%; p<0.05 of group A and group B respectively).

• 한국가축번식학회지
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• 제15권2호
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• pp.103-107
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• 1991

This study was carried out to investigate the survival rate in vitro culture after frozen-thawed to used DMSO(dimethyl sulfoxide), glycerol and ethylene glycol of cryoprotective agents at the zona pellucida removed and encased into alien bisected embryo of the mouse early embryos. The results obtained from this study were as follows : 1. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the zona pellucida removed bisected morula was 46.6%, 35.8% and 27.3%, total or mean were 36.6%, respectively. 2. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the encased into alien bisected morula was 70.6%, 65.3% and 66.4%, total or mean were 67.4%, respectively. 3. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the zona pellucida removed bisected blastocysts was 50.4%, 36.7% and 30.4%, total of mean were 39.2%, respectively. 4. The survival rate of in vitro culture after frozen-thawed to used cryoprotective agents of three kinds at the encased into alien bisected blastocysts was 71.1%, 66.7% and 63.9%, total or mean were 67.2%, respectively.

• 한국수정란이식학회지
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• 제11권1호
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• pp.27-33
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• 1996

The efficient cryopreservation of embryos requires optimal times of dehydration and rehydration This study was carried out to investigate the effect of various times of dehydration and rehydration The effects were evaluated through testing morphological normality and developmental ability of 1 cell mouse embryos which were ultrarapidly frozen and thawed. The 1 cell embryos were dehydrated for 1.5, 3, 5, and 10 minutes using mPBS-BSA containing 3.SM DMSO and 0.25M sucrose on cooling chamber or on ice. After ultrarapidly frozen and thawed, they were rehydrated for 0, 0.5 and 5 minutes with mPBS-BSA containing 0.25M sucrose at room temperature. The results obtained were as follows: The embryos that were rehydrated for 0.5 minutes showed higher normality than the embryos for 0 and 5 minutes did. The embryos that were dehydrated for 10 minutes showed higher normality than the embryos for 1.5, 3, and 5 minutes did. The developmental ability of normal thawed-embryos was high in 10 minute dehydration treatment compared to other treatments. However, it was not affected by cooling methods (on ice and on cooling chamber) for embryo dehydration.