骨髓间充质干细胞亚群心肌分化基因的筛选  

Screening of myocardial differentiation genes from bone marrow mesenchymal stem cells subpopulations

作　　者：滕小梅[1] 贺继刚[1] 王霄琳[1] 吴伟华[2] 

机构地区：[1]苏州大学附属第一医院心血管外科,江苏苏州215006 [2]苏州大学附属第一医院检验科,江苏苏州215006

出　　处：《西南国防医药》2014年第10期1062-1065,共4页Medical Journal of National Defending Forces in Southwest China

基　　金：苏州市科技计划项目(SYS201327)

摘　　要：目的利用基因芯片筛选小鼠骨髓间充质干细胞4个亚群的差异表达基因,寻找心肌分化特异性基因。方法将小鼠骨髓间充质干细胞通过CD45和CD31的磁珠分选,获得SCA-1^＋/CD45^-/CD31^-、SCA-1^＋/CD45^-/CD31^＋、SCA-1^＋/CD45^＋/CD31^＋及SCA-1^＋/CD45^＋/CD31^-4群细胞。采用基因芯片技术完成Agilent小鼠全基因4＊44K芯片表达谱基因检测,分析差异表达基因。根据基因芯片中所检测的基因表达量在SCA-1^＋/CD45^＋/CD31^＋亚群中最高,且为其他亚群表达量的两倍以上为原则,通过RT-PCR验证各亚群及未分选细胞中的差异基因的表达量。结果根据皮尔森关联算法计算4组细胞亚群之间的关联值位于0.979～0.995之间,通过RT-PCR检测发现,Rock2基因在SCA-1^＋CD45^-CD31^-亚群中表达最高,而在SCA-1^＋CD45^＋CD31^＋亚群中表达很低,Itga4在未分选群中表达最高;Cxadr在SCA-1^＋CD45^-CD31^-亚群中表达最高;Epor在SCA-1^＋CD45^-CD31^＋亚群中表达最高;myl3在SCA-1^＋CD45^＋CD31^＋亚群中表达最高。结论 Rock2、Cxadr、Itga4、Epor和myl3基因在干细胞亚群向心肌分化中发挥不同作用。Objective To find specific myocardial differentiation genes by using gene chip to screen the differentially expressed genes from 4 bone marrow mesenchymal stem cells subpopulation of mice.Methods The bone marrow mesenchymal stem cells subpopulations of mice were sorted by means of magnetic beads of CD45 and CD31,and four cells subpopulations were obtained,i.e.,SCA-1^＋/CD45^-/CD31^-,SCA-1^＋/CD45^-/CD31^＋,SCA-1^＋/CD45^＋/CD31^＋and SCA-1^＋/CD45^＋/CD31^-.And then,the gene chip technology was used to complete the detection of Agilent mice＇whole 4＊ 44 K gene expression and the differentially expressed genes were analyzed.Based on the principle that the expression level of the genes shall be the highest in the subpopulation of SCA-1^＋/CD45^＋/CD31^＋and it shall be 2 times of that in other subpopulations,the expression levels of the differentially expressed genes of the subpopulations and other unsorted subpopulation were determined by means of RT-PCR.Results According to the results of Pearson correlation algorithm,the correlation value of the 4 cells subpopulations was between 0.979 and 0.995;according to the RT-PCR detection,the expressed gene number of Rock2 was the highest in SCA-1^＋CD45^-CD31^-,but the lowest in SCA-1^＋CD45^＋CD31^＋,while that of Itga4 was the highest in the unsorted subpopulation.Besides,the expressed gene number of Cxadr was the highest in SCA-1^＋CD45^-CD31^-,that of myl3 was the highest in SCA-1^＋CD45^＋CD31^＋.Conclusion The genes Rock2,Cxadr,Itga4,and Epor and myl3 play different roles in the differentiated course from the bone marrow mesenchymal stem cells subpopulations to myocardial cells.

关 键 词：骨髓 间充质干细胞 心肌分化 基因芯片 

分 类 号：R457.7[医药卫生—治疗学]