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말초신경재생을 위한 hNGF-$eta$ recombinant Adenovirus의 제작 및 수종세포주에서 신경성장인자의 발현
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  • 말초신경재생을 위한 hNGF-$eta$ recombinant Adenovirus의 제작 및 수종세포주에서 신경성장인자의 발현
저자명
고은봉,정헌종,안강민,김윤태,박희정,성미애,김남열,유상배,명훈,황순정,김명진,김성민,장정원,이종호,Gao. En-Feng,Chung. Hun-Jong,Ahn. Kang-Min,Kim. Yoon-Tae,Park. Hee-Jung,S
간행물명
대한악안면성형재건외과학회지
권/호정보
2005년|27권 5호|pp.446-456 (11 pages)
발행정보
대한악안면성형재건외과학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

Nerve growth factor(NGF) has a critical role in peripheral nerve regeneration. The aim of this study is to construct a well-functioning hNGF-$eta$ recombinat adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with adenovirus mediated hNGF-$eta$ gene transfection into Schwann cells. First PCR associated cloning of GFP-tagged hNGF-$eta$ which was ligated into E1/E3 deleted adenoviral vector was performed and tranfected into E. coli to construct hNGF-$eta$ recombinant adenovirus. After production of recombinat adenovirus in a large scale, its transfection efficiency, expression, and function were evaluated using cell lines or primarily cultured cells of HEK293 cells, Schwann cells, fibroblast(NIH3T3) and myocyte(CRH cells). GFP expression was observed in 90% of infected cells compared to uninfected cells. Total mRNA isolated from hNGF-$eta$ recombinat adenoviru infected cells showed strong RT-PCR band, however, LacZ recombinant adenovirus infected or uninfected cells did not. NGF quantification by ELISA showed a maximal release of 18.865 +/- 0.31ng/mL at 4th day. PC-12 cells exposed to media with hNGF-$eta$ recombinant adenovirus infected Schwann cell demonstrated higher levels of differentiation compared with controls. We generated hNGF-$eta$ recombinant adenovirus and induced over expression of NGF successfully in nonneuronal and neuronal cells. Following these result, it is expected to develop an improved treatment strategy peripheral nerve regeneration using the hNGF-$eta$ gene transfected cells.