• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.188-188
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• 2004

근래에 많은 형질전환 실험동물이 생산되어 인간을 대체하여 전임상실험에 사용되고 있으며, 그 요구는 점차 증가되고 있다. 그러나 지금까지 생산된 것은 소형 실험동물에 국한되고 있어, 사람과의 그 크기와 기능에 거리가 있었던 것이 사실이다. 본 실험은 중형 실험동물이자 사람과 여러 기관 및 조직이 유사하다고 알려진 돼지로부터 난자를 채취하여 난자의 핵을 GFP가 발현된 세포의 핵으로 치환하여 태어난 산자를 재복제하여 생산된 돼지 신생자를 이용하여 각 기관 및 조직에서 GFP의 발현을 RT-PCR, Northern blot 및 immunocytochemistry 방법으로 분석하였다. (중략)

• Reproductive and Developmental Biology
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• v.35 no.1
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• pp.23-31
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• 2011

Two piglets and one juvenile pig were used to investigate closely what types of cells express green fluorescent protein (GFP) and if any, whether the GFP-tagged cells could be used for stem cell transplantation research as a middle-sized animal model in bone marrow cells of recloned GFP pigs. Bone marrow cells were recovered from the tibia, and further analyzed with various cell lineage markers to determine which cell lineage is concurrently expressing visible GFP in each individual animal. In the three animals, visible GFP were observed only in proportions of the plated cells immediately after collection, showing 41, 2 and 91% of bone marrow cells in clones #1, 2 and 3, respectively. The intensity of the visible GFP expression was variable even in an individual clone depending on cell sizes and types. The overall intensities of GFP expression were also different among the individual clones from very weak, weak to strong. Upon culture for 14 days in vitro (14DIV), some cell types showed intensive GFP expression throughout the cells; in particular, in cytoskeletons and the nucleus, on the other hand. Others are shown to be diffused GFP expression patterns only in the cytoplasm. Finally, characterization of stem cell lineage markers was carried out only in the clone #3 who showed intensive GFP expression. SSEA-1, SSEA-3, CD34, nestin and GFAP were expressed in proportions of the GFP expressing cells, but not all of them, suggesting that GFP expression occur in various cell lineages. These results indicate that targeted insertion of GFP gene should be pursued as in mouse approach to be useful for stem cell research. Furthermore, cell- or tissue-specific promoter should also be used if GFP pig is going to be meaningful for a model for stem cell transplantation.