• Reproductive and Developmental Biology
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• v.31 no.1
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• pp.29-33
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• 2007

This study was conducted to examine the development of in vitro fertilized (IVF) and nuclear transfer (NT) embryos following vitrification IVF and NT embryos developed to the blastocyst stage were equilibrated by 3 steps, vitrified and thawed, and their survival and hatching rates were examined. In IVF embryos, higher survival (82.1%, 96/117) and hatching rates (64.1%, 75/117) were obtained respectively after thawing and culture in expanded blastocysts compared to blastocysts (p<0.05). High survival and hatching rates were also obtained by vitrification of NT blastocysts, especially in expanded and hatching blastocysts (81.1 and 78.3%, respectively). The result of this study shows that IVF and NT blastocysts, especially late stage blastocysts, are successfully cryopreserved by vitrification.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2000.11a
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• pp.5-20
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• 2000

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.42-42
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• 2003

복제기술은 기존의 형질전환 동물 생산의 효율을 향상시킬 수 있는 기술로서 인정하고 있으며 또한 이를 이용하여 형질전환 동물의 생산이 이루어지고 있다. 따라서 본 연구는 표지유전자 (GFP)와 유용유전자 (hFSH)를 이용하여 임신 45일령에 채취한 태아섬유아세포에 transfection 하고, transfection 된 세포의 효율적인 선발과 이를 이용한 형질전환 복제 수정란을 생산하고자 실시하였다. 대조구 (KbFF), GFP (79KbFF-GFP c-3) 및 hFSH (79KbFF-hFSH n-1)에 공시한 세포는 모두 동일한 태아유래의 세포 (모 79, 부 KPN178,♂)를 이용하였다. pAB-eGFP와 hFSH 유전자는 각파 electroporation 방법을 이용하여 transfection 하고, 이를 2주 동안 G418로 배양하며 selection 하였다.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.135-135
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• 2003

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.419-424
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• 1998

The present study was carried out to examine the efficiency of cloning of transgenic embryos by nuclear transfer(NT) using gene-injected rabbit embryos. The rabbit embryos at pronuclear stage were microinjected with methallothionein-human growth hormone(MT-hGH) gene and cultured to 8- and 16-cell in TCM-199 containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% $CO_2$incubator. The recipient oocytes were collected from the oviducts 14~16 h after hCG injection. The oocytes were enucleated and activated with 5$\mu$M ionomycin and 2mM 6-dimethylaminopurine. Blastomeres form gene-injected embryos were transferred into the enucleated oocytes by micromanipulation. The nuclear transplant oocytes were electrofused and co-cultured with rabbit oviductal cells. Following 120 h of culture, blastocysts were prepared for gene analysis by polymerase chain reaction(PCR). In previous experiment, the rate of gene-positive embryos detected by the nested PCR analysis was significantly decreased while developing to blastocyst(25%)(Kang et al., 1998). The fusion rate of gene-injected blastomeres was significantly(P＜0.05) lower than non-injected blastomeres(66% vs 80%). However, the NT embryos that were derived from gene-injected donor embryos did not differ from control embryos in development to the blastocyst stage(39% vs 31%). Of the 43 NT blastocysts developed from the gene-injected donor embryos, twelve(28%) were positive for the injected DNA. The results indicate that NT with gene-injected embryos can be successfully used for cloning and multiplication of transgenic embryos, furthermore applicable to improvement of transgenic animal production.

• Journal of Veterinary Clinics
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• v.24 no.3
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• pp.295-299
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• 2007

The detrimental effects of gene transfection on embryo development and the molecular mechanism behind the differential expression of genes related to early embryo development were assessed in the production of transgenic cow embryos through somatic cell nuclear transfer (NT). Parthenogenetic, IVF, and transgenic NT embryos derived from ${\alpha}_1$-antitrypsin transfected ear fibroblast cells was produced. To investigate the molecular mechanism behind lower developmental competence of transgenic NT embryos, the differential mRNA expression of three genes ($IFN-{\tau}$, Oct4, Fgf4) in the 3 types of embryo (Parthenogenetic, IVF, transgenic NT) was examined. RNA was extracted from ten blastocysts derived from 3 types of embryos and reverse-transcripted for synthesis of the first cDNA. The quantification of 3 gene transcripts ($IFN-{\tau}$, Oct4, and Fgf4) was carried out in three replicate by quantitative real-time reverse transcriptase PCR. Expression level of $IFN-{\tau}$ mRNA was significantly higher in transgenic NT embryos than parthenogenetic and IVF embryos (P<0.05). However, expression level of Oct4 and Fgf4 of transgenic NT embryos was significantly lower than IVF embryos (P<0.05). Altered levels of these three mRNA transcripts may explain some of the embryonic/fetal/neonatal abnormalities observed in offspring from transgenic NT embryos.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.05a
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• pp.37-42
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• 2004

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2000.11a
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• pp.34-45
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• 2000

• Journal of the korean veterinary medical association
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• v.37 no.10
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• pp.938-948
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• 2001

최근 대동물임상분야에서도 소의 번식장애뿐만 아니라 첨단번식기술에 대한 연구가 지속적으로 이뤄져오고 있으며, 관련분야에 종사하는 임상가, 연구자들에게도 수정란이식, 체세포복제수정란이식분야 등에 대한 연구가 시급하게 요구되고 있는 실정이다. 따라서 필자는 대동물임상가들과 관련연구자에게 도움이 되었으면 하는 바램을 피력하면서 지난 수의사회지 8월호에 이어<일본 북해도 삿뽀로시 에베츠에 소재하는 낙농학원대학(酪農學園大學:Rakuno Gakuen University)에서 지난 5월 26일(토) 오전 8시 50분부터 오후 6시까지 대동물임상수의사들을 대상으로 한 교육세미나>에서 발표된 자료중 소개를 하지 않은 부분을 다시금 시간의 여유를 내어 번역을 하여 소개하고자 한다. 지난 8월호에 이어 필자가 일본어로 된 자료를 번역하는 과정에서 우리말로 표현하는데 무리가 있는 부분은 알기쉽게 평이하게 정리하였음을 서두에 밝혀 둔다. 또한 우리 임상수의사들도 최근 관심이 증대되고 있는 수정란이식과 체세포복제수정란 등에 대한 전문지식과 연구정보를 입수, 습득하여 나가지 않으면 아니된다 하겠다.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.42-42
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• 2002

본 연구는 한국 재래산양의 핵이식에 있어서 공여핵은 귀세포를, 수핵난자는 소 및 돼지의 난포란을 이용하여 핵이식을 실시하여 복제수정란의 체외발달율과 이종간의 핵이식 가능성을 검토하였다. 공여핵은 재래산양의 귀세포를 채취하여 10% FBS 가 첨가된 TCM-199 배양액으로 체외 배양을 실시하여 monolayar Confluent 형성후 0.25% Trypsin-EDTA을 처리하여 계대배양을 실시하였다. (중략)