中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
37期
5916-5922
,共7页
王晓%甄雷%缪黄泰%吴星欣%任红梅%师树田%乔岩%刘新民%阙斌%聂绍平
王曉%甄雷%繆黃泰%吳星訢%任紅梅%師樹田%喬巖%劉新民%闕斌%聶紹平
왕효%견뢰%무황태%오성흔%임홍매%사수전%교암%류신민%궐빈%섭소평
干细胞%骨髓干细胞%冠状静脉逆行灌注%骨髓间充质干细胞%细胞移植%碱性成纤维细胞生长因子%分化%急性心肌梗死%国家自然科学基金
榦細胞%骨髓榦細胞%冠狀靜脈逆行灌註%骨髓間充質榦細胞%細胞移植%堿性成纖維細胞生長因子%分化%急性心肌梗死%國傢自然科學基金
간세포%골수간세포%관상정맥역행관주%골수간충질간세포%세포이식%감성성섬유세포생장인자%분화%급성심기경사%국가자연과학기금
bone marrow%mesenchymal stem celltransplantation%fibroblast growth factor 2%myocardial infarction
背景：体外研究显示，碱性成纤维细胞生长因子能促进骨髓间充质干细胞向心肌样细胞分化。然而，在体内环境下经冠状静脉逆行灌注碱性成纤维细胞生长因子能否促进骨髓间充质干细胞分化尚不明确。目的：探讨经冠状静脉逆行灌注碱性成纤维细胞生长因子对骨髓间充质干细胞在体内分化的影响。方法：①杂种犬12只，采用密度梯度离心与贴壁培养法在体外分离、培养骨髓间充质干细胞，增强型绿色荧光蛋白慢病毒载体转染骨髓间充质干细胞并分析转染率。②开胸结扎法建立急性心肌梗死模型，1周后将存活犬(n=10)随机分为骨髓间充质干细胞组(n=5)和碱性成纤维细胞生长因子+骨髓间充质干细胞组(n=5)，采用OTW球囊经冠状静脉逆行灌注碱性成纤维细胞生长因子和增强型绿色荧光蛋白标记的骨髓间充质干细胞。移植后1周，免疫荧光法比较两组梗死心肌内增强型绿色荧光蛋白阳性细胞共表达Ⅷ因子和肌钙蛋白I的数量。结果与结论：增强型绿色荧光蛋白慢病毒成功转染骨髓间充质干细胞，转染率达85%；灌注后免疫荧光显示，23.5%的切片可以看到增强型绿色荧光蛋白标记的骨髓间充质干细胞阳性细胞；碱性成纤维细胞生长因子+骨髓间充质干细胞组增强型绿色荧光蛋白共表达Ⅷ因子和肌钙蛋白 I 的细胞数量明显高于骨髓间充质干细胞组(P<0.05)。因此，经冠状静脉逆行灌注碱性成纤维细胞生长因子能更有效地促进骨髓间充质干细胞分化成血管内皮细胞和心肌细胞；采用该途径联合灌注碱性成纤维细胞生长因子和骨髓间充质干细胞有望发挥协同作用，更好地促进心脏修复。
揹景：體外研究顯示，堿性成纖維細胞生長因子能促進骨髓間充質榦細胞嚮心肌樣細胞分化。然而，在體內環境下經冠狀靜脈逆行灌註堿性成纖維細胞生長因子能否促進骨髓間充質榦細胞分化尚不明確。目的：探討經冠狀靜脈逆行灌註堿性成纖維細胞生長因子對骨髓間充質榦細胞在體內分化的影響。方法：①雜種犬12隻，採用密度梯度離心與貼壁培養法在體外分離、培養骨髓間充質榦細胞，增彊型綠色熒光蛋白慢病毒載體轉染骨髓間充質榦細胞併分析轉染率。②開胸結扎法建立急性心肌梗死模型，1週後將存活犬(n=10)隨機分為骨髓間充質榦細胞組(n=5)和堿性成纖維細胞生長因子+骨髓間充質榦細胞組(n=5)，採用OTW毬囊經冠狀靜脈逆行灌註堿性成纖維細胞生長因子和增彊型綠色熒光蛋白標記的骨髓間充質榦細胞。移植後1週，免疫熒光法比較兩組梗死心肌內增彊型綠色熒光蛋白暘性細胞共錶達Ⅷ因子和肌鈣蛋白I的數量。結果與結論：增彊型綠色熒光蛋白慢病毒成功轉染骨髓間充質榦細胞，轉染率達85%；灌註後免疫熒光顯示，23.5%的切片可以看到增彊型綠色熒光蛋白標記的骨髓間充質榦細胞暘性細胞；堿性成纖維細胞生長因子+骨髓間充質榦細胞組增彊型綠色熒光蛋白共錶達Ⅷ因子和肌鈣蛋白 I 的細胞數量明顯高于骨髓間充質榦細胞組(P<0.05)。因此，經冠狀靜脈逆行灌註堿性成纖維細胞生長因子能更有效地促進骨髓間充質榦細胞分化成血管內皮細胞和心肌細胞；採用該途徑聯閤灌註堿性成纖維細胞生長因子和骨髓間充質榦細胞有望髮揮協同作用，更好地促進心髒脩複。
배경：체외연구현시，감성성섬유세포생장인자능촉진골수간충질간세포향심기양세포분화。연이，재체내배경하경관상정맥역행관주감성성섬유세포생장인자능부촉진골수간충질간세포분화상불명학。목적：탐토경관상정맥역행관주감성성섬유세포생장인자대골수간충질간세포재체내분화적영향。방법：①잡충견12지，채용밀도제도리심여첩벽배양법재체외분리、배양골수간충질간세포，증강형록색형광단백만병독재체전염골수간충질간세포병분석전염솔。②개흉결찰법건립급성심기경사모형，1주후장존활견(n=10)수궤분위골수간충질간세포조(n=5)화감성성섬유세포생장인자+골수간충질간세포조(n=5)，채용OTW구낭경관상정맥역행관주감성성섬유세포생장인자화증강형록색형광단백표기적골수간충질간세포。이식후1주，면역형광법비교량조경사심기내증강형록색형광단백양성세포공표체Ⅷ인자화기개단백I적수량。결과여결론：증강형록색형광단백만병독성공전염골수간충질간세포，전염솔체85%；관주후면역형광현시，23.5%적절편가이간도증강형록색형광단백표기적골수간충질간세포양성세포；감성성섬유세포생장인자+골수간충질간세포조증강형록색형광단백공표체Ⅷ인자화기개단백 I 적세포수량명현고우골수간충질간세포조(P<0.05)。인차，경관상정맥역행관주감성성섬유세포생장인자능경유효지촉진골수간충질간세포분화성혈관내피세포화심기세포；채용해도경연합관주감성성섬유세포생장인자화골수간충질간세포유망발휘협동작용，경호지촉진심장수복。
BACKGROUND:In vitro studies have demonstrated that basic fibroblast growth factor (bFGF) promote the differentiation of bone marrow mesenchymal stem cells (BMSCs) into cardiomyocyte-like cells. However, it is unclear whether coronary venous retroperfusion of bFGF stimulates BMSCs differentiation in vivo. OBJECTIVE:To evaluate the effects of coronary venous retroperfusion of bFGF on BMSCs differentiation in vivo. METHODS:BMSCs from 12 dogs were isolated by density gradient centrifugation and expanded in vitro. These cells were transfected by enhanced green fluorescence protein (EGFP) lentiviral vector and the transfection efficiency was analyzed. Acute myocardial infarction was induced by ligation of left anterior descending coronary artery. After 1 week, 10 survival animals were randomized to BMSCs group (n=5) and bFGF+BMSCs group (n=5). bFGF-and EGFP-positive BMSCs were reversely infused via coronary vein using over-the-wire bal oon catheter. One week after infusion, the number of EGFP-positive cells co-staining factor VIII and troponin I was compared between the two groups by immunofluorescence method. RESULTS AND CONCLUSION:BMSCs were successful y transfected by EGFP and the transfection efficiency was 85%. Immunofluorescence showed that EGFP-positive BMSCs were observed in 23.5%of slides. There were more EGFP-positive cells co-staining VIII and troponin I in the bFGF+BMSCs group than in the BMSCs group (P<0.05). Thus, the coronary venous retroperfusion of bFGF enhances the differentiation of BMSCs into vascular endothelial cells and cardiomyocytes. Combined delivery of bFGF and BMSCs can exert a synergistic effect to promote cardiac repair.
