富含半乳糖凝集素-1基因修饰的骨髓间充质干细胞与脱细胞真皮基质补片共培养的研究  

The experiment study of mesenchymal stem cells modified wtih galectin-1 gene co-cultured with dermal matrix patch

作　　者：刘先桂[1] 张廷涛[1] 张秋雷[1] 刘韦成[1] 丁召[1] 钱群[1] 江从庆[1] 

机构地区：[1]武汉大学中南医院结直肠肛门外科湖北省肠病医学临床研究中心肠病湖北省重点实验室湖北省卫生和计划生育委员会结直肠肛门外科质量控制中心,430071

出　　处：《中华实验外科杂志》2015年第5期1048-1050,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81070387）

摘　　要：目的 探讨半乳糖凝集素-1（Galectin-1）基因修饰的骨髓间充质干细胞（BMSCs）与脱细胞真皮基质（ADM）补片共培养的可行性.方法 将Ad-绿色荧光蛋白（GFP）-Galectin-1转染间充质干细胞（MSCs）后,荧光显微镜下观察荧光表达,通过实时定量反转录聚合酶联反应（RT-qPCR）法检测Galectin-1基因表达,酶联免疫吸附试验（ELISA）法检测Galectin-1蛋白表达,Western blot检测肌间线蛋白（Desmin）表达,共聚焦荧光显微镜下观察Ad-GFP-Galectin-1转染BMSCs与ADM补片共生长情况.结果 Ad-GFP-Galectin-1转染BMSCs后,荧光显微镜下可见绿色荧光,72 h达高峰;RT-qPCR检测Galectin-1 mRNA的表达,Galectin-1 mRNA在空白对照组（PBS组）中相对表达水平为1.00;在Ad-GFP-Galectin-1（Ad-GFP-Galectin-1组）和Ad-GFP（Ad-GFP组）转染BMSCs中相对表达水平分别为4.11和1.09;ELISA证实BMSCs表达Galectin-1、Galectin-1表达量在Ad-GFP-Galectin-1组、Ad-GFP组和PBS组中分别为48 h：（197.28±8.78）、（25.52±1.56）、（24.53±2.96） μg/L;72 h：（264.33±5.43）、（25.40±2.06）、（25.46±1.13） μg/L;96 h：（172.41±3.61）、（25.22±1.61）、（28.01±1.26） μg/L;120 h：（155.53±7.25）、（24.84±2.35）、（26.65±1.54） μg/L. Western blot证实大鼠Desmin表达升高;共聚焦荧光显微镜下观察Ad-GFP-Galectin-1转染的BMSCs与ADM融合生长良好.结论 Galectin-1能诱导BMSCs合成Desmin;Ad-GFP-Galectin-1转染的BMSCs与脱细胞真皮基质（ADM）补片融合生长良好.Objective To explore the feasibihty of mesenchymal stem cells （MSCs） modified wtih galectin-1 gene co-cultured with dermal matrix patch.Methods After the bone marrow MSCs （BMSCs） were transfected with Ad-green fluorescent protein （GFP）-galectin-1,the fluorescence in BMSCs was observed under fluorescence microscope.Real-time quantitative reverse transcriptase-polymerase chain reaction（RT-qPCR） was used to detect the expression of galectin-1 gene,and the protein expression of galectin-1 and desmin was detected by enzyme linked immune sorbent assay （ELISA） and Western blotting,respectively.The growth of BMSCs transfected with Ad-GFP-galectin-1 and co-cultured with acellular dermal matrix patch （ADM） was observed.Results The green fluoresence was observed and reached to peak at 72 h after the transfection of BMSCs with Ad-GFP-galectin-1 under fluorescence microscope.The result of RT-qPCR and ELISA confirmed that BMSCs over-expressed galectin-1.The RT-qPCR result of galectin-1,which the relative expression folds was 1.00 in phosphate buffer （PBS） group,4.11 in Ad-GFP-galectin-1 group,1.09 in Ad-GFP group;The result of galectin-1 measured by ELISA was that 48 h：（197.28 ± 8.78）,（25.52 ± 1.56）,（24.53 ± 2.96） μg/L;72 h：（264.33 ±5.43）,（25.40±2.06）,（25.46±1.13） μg/L;96h：（172.41 ±3.61）,（25.22±1.61）,（28.01±1.26） μg/L;120 h：（155.53 ±7.25）,（24.84 ±2.35）,（26.65 ± 1.54） μg/L ordered by Ad-GFP-galectin-1 group,Ad-GFP group and PBS group.The expression of desmin which co-cultured with dermal matrix patch （ADM） were well grown.Conclusion Galectin-1 could induce BMSCs to express desmin.BMSCs transfected with Ad-GFP-galectin-1 showed good compatibihty with ADM.

关 键 词：半乳糖凝集素-1 骨髓间充质干细胞 脱细胞真皮基质 共培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]