CAJ | 학술논문

目的：探讨培养大鼠骨髓间充质干细胞（BMSCs）细胞株、表型鉴定及利用重组质粒（pEGFP/Ang-1）转染大鼠 BMSCs细胞株的实验方法。方法采用贴壁筛选法和密度梯度离心法获取BMSCs，纯化大鼠BMSCs后，以重组质粒pEGFP/Ang-1转染，G 418筛挑阳性克隆，倒置荧光显微镜下观察增强型绿色荧光蛋白的表达，检测Ang-1表达量的时间变化。结果采用贴壁筛选法和密度梯度离心法成功获取BMSCs细胞株，成功将血管生成素-1重组质粒（pEGFP/Ang-1）转染大鼠BMSCs，3 d可稳定表达Ang-1，7 d以后开始减弱。结论成功运用贴壁筛选法和密度梯度离心法分离培养的大鼠BMSCs，并重组质粒pEGFP/Ang-1转染大鼠BMSCs可有效表达。
목적：탐토배양대서골수간충질간세포（BMSCs）세포주、표형감정급이용중조질립（pEGFP/Ang-1）전염대서 BMSCs세포주적실험방법。방법채용첩벽사선법화밀도제도리심법획취BMSCs，순화대서BMSCs후，이중조질립pEGFP/Ang-1전염，G 418사도양성극륭，도치형광현미경하관찰증강형록색형광단백적표체，검측Ang-1표체량적시간변화。결과채용첩벽사선법화밀도제도리심법성공획취BMSCs세포주，성공장혈관생성소-1중조질립（pEGFP/Ang-1）전염대서BMSCs，3 d가은정표체Ang-1，7 d이후개시감약。결론성공운용첩벽사선법화밀도제도리심법분리배양적대서BMSCs，병중조질립pEGFP/Ang-1전염대서BMSCs가유효표체。
Objective To train the successful bone marrow mesenchymal stem cells ( BMSCs ) and phenotypic characterization , and investigate the beneficial effects of recombinant plasmid ( pEGFP/Ang-1) transfected rats BMSCs .Methods Using adherence screening and density gradient centrifugation methods , purified rats BMSCs were obtained .After transfection with pEGFP/Ang-1, the BMSCs were screened with G418, and the positive clones were sequentially confirmed by expression of enhanced green fluorescent protein (GFP) with fluorescence microscope, and expression of angiogenin-1(Ang-1) was tested by Western blot.Results Using adherence screening and density gradient centrifugation methods , purified rats BMSCs were successfully obtained .The successfully stable transfection of pEGFP-N1-Ang-1 in BMSCs had be stably expressed Ang-1 3 days,later started to diminish.Conclusions The successful use of adherent culture method and density gra-dient centrifugation of rat BMSCs and the recombinant plasmid pEGFP /Ang-1 being transfected into the rat BMSCs could effectively express .