CAJ | 학술논문

背景:近些年来应用相关干细胞及生长因子促进肺再生修复肺气肿病变的研究逐渐成为热点,那么通过干细胞携带相关外源生长因子基因能否更好的修复肺气肿病变值得关注.目的:观察携带绿色荧光蛋白基因的腺病毒(adenovirus vecto rmediated green fluorescence protein,Ad-GFP)转染骨髓间充质干细胞的效率、对细胞增殖的影响,及携带目的基因的间充质于细胞尾静脉移植在肺气肿大鼠肺部的定居情况.方法:采用密度梯度离心结合贴壁培养法分离、培养间充质干细胞,流式细胞仪鉴定;在不同感染复数时,激光共聚焦显微镜检测转染效果及转染效率,MTT法检测转染48 h的细胞活性:Wistar大鼠16只,随机分为模型组和对照组,每组8只,采用单纯被动烟熏法制备肺气肿大鼠模型.通过尾静脉移植Ad-GFP修饰的间充质干细胞,于24 h后取大鼠肺组织做快速冰冻切片,在荧光显微镜下观察间充质干细胞在肺部的分布情况.结果与结论:实验成功获得大鼠间充质干细胞,Ad-GFP成功转染间充质干细胞,且在MOI=200时,转染效率达到88.42%.在不同MOI下,Ad-GFP转染间充质干细胞对细胞增殖无影响(P＞0.05).移植24 h后,模型组大鼠及对照组大鼠肺部可见到发绿色荧光的间充质干细胞,且模型组荧光强度明显强于对照组(P＜0.05).提示导入目的基因可能不会影响骨髓间充质干细胞增殖和归巢特性.
배경:근사년래응용상관간세포급생장인자촉진폐재생수복폐기종병변적연구축점성위열점,나요통과간세포휴대상관외원생장인자기인능부경호적수복폐기종병변치득관주.목적:관찰휴대록색형광단백기인적선병독(adenovirus vecto rmediated green fluorescence protein,Ad-GFP)전염골수간충질간세포적효솔、대세포증식적영향,급휴대목적기인적간충질우세포미정맥이식재폐기종대서폐부적정거정황.방법:채용밀도제도리심결합첩벽배양법분리、배양간충질간세포,류식세포의감정;재불동감염복수시,격광공취초현미경검측전염효과급전염효솔,MTT법검측전염48 h적세포활성:Wistar대서16지,수궤분위모형조화대조조,매조8지,채용단순피동연훈법제비폐기종대서모형.통과미정맥이식Ad-GFP수식적간충질간세포,우24 h후취대서폐조직주쾌속빙동절편,재형광현미경하관찰간충질간세포재폐부적분포정황.결과여결론:실험성공획득대서간충질간세포,Ad-GFP성공전염간충질간세포,차재MOI=200시,전염효솔체도88.42%.재불동MOI하,Ad-GFP전염간충질간세포대세포증식무영향(P＞0.05).이식24 h후,모형조대서급대조조대서폐부가견도발록색형광적간충질간세포,차모형조형광강도명현강우대조조(P＜0.05).제시도입목적기인가능불회영향골수간충질간세포증식화귀소특성.
BACKGROUND:Recently,application of stem cells and growth factor to promoting lung regeneration in repair of emphysema lesion has been a hot focus in study.Thus,it is worth to pay attention on whether stem cells carrying relevant foreign growth factor gene can repair emphysema lesion.OBJECTIVE:To evaluate the effidency of adenovirus vector mediated green fluorescence protein(Ad-GFP)transfecting bone marrow mesenchymal stem cells(BMSCs)and its effect on the cell proliferation,to explore oriented migration of intravenously administrated BMSCs transfected with Ad-GFP in the lung tissues of pulmonary emphysema rats.METHODS:MSCs were separated and purified from the bone marrow of rats by density gradient centrifugation and by adherence.At different multiplicity of infection(MOI),transfection efficiency was observed by laser confocal microscopy.At 48 hours of transfection,MTT method was used to evaluate the proliferation of MSCs.A total of 16 Wistar rats were randomly divided into emphysema model group and control group(n=8).Model rats were established by exposure to cigarette smoke.MSCs,transfected with Ad-GFP,were grafted into the body of rats via tail vein.Lungs derived at 24 hours after implantation,and frozen sections were made.Migration and survival of MSCs in the lung tissues were observed by fluorescence microscopy.RESULTS AND CONCLUSION:MSCs from Wistar rats were successfully cultured,grew well and infected by Ad-GFP.The highest transfection effincincy(88.42 %)could be achieved at MOI of 200.Green fluorescent protein labeling had little effect on proliferation of MSCs by different MOI(P＞0.05).At 24 hours posttransplantation,the green fluorescence-positive tissue was Found in the lung tissues of emphysema model group and control group.Compared with control group,the expression of GFP in lung tissues was higher in emphysema model group(P＜0.05).These suggested that introduction of target gene cannot affect proliferation and homing property of BMSCs.