目的:探索构建稳定表达IL-33的原代大鼠骨髓间充质干细胞(BMSC)细胞系的方法。方法:体外分离、培养、鉴定大鼠骨髓间充质干细胞,并进行ELISA实验检测转染前IL-33的表达情况。构建GV303 Rat IL-33慢病毒载体,并进行慢病毒的包装。3d后荧光显微镜下观察GFP的表达,分别使用Western blot、ELISA检测转染后的BMSC中IL-33的表达。结果:大鼠BMSC被成功分离并传代,流式细胞术鉴定BMSC可得到CD90、CD29为阳性,CD11b、CD45、CD34为阴性,与之前的文献报道相符。并且BMSC被成功诱导分化为骨细胞、软骨细胞、脂肪细胞。对转染前的BMSC行ELISA实验检测IL-33的表达,结果为阴性。病毒转染3d后于荧光显微镜下可观察到GFP的表达;ELISA实验检测转染后BMSC中IL-33的表达,结果为阳性;Western blot亦可以检测到IL-33的表达。结论:应用GV303慢病毒载体可以建立稳定表达IL-33的大鼠BMSC原代细胞系。
【英文摘要】
AIM:To explore a way to establish a rat bone mesenchymal stem cell(BMSC) line expressing Interleukin(IL)-33. METHODS:BMSCs were isolated from 6-week-old Wistar Rat.GV303 virus were used as a IL-33 gene carrier to tansfect isolated BMSCs.Three days later,transfected BMSCs were observed under a fluorescence microscope.The expression level of IL-33 of transfected BMSCs was detected by Western blot and ELISA. RESULTS:The BMSCs were successfully isolated,since flow cytometry results showed that rat BMSCs CD90 an...
| 
推荐 
点击下列相关研究机构和相关文献作者，可以直接查到这些机构和作者被《中国知识资源总库》收录的其它文献，使您全面了解该机构和该作者的研究动态和历史。
