CAJ | 학술논문

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垣/ CD45原/ CD31原、SCA-1垣/ CD45原/ CD31垣、SCA-1垣/CD45垣/ CD31垣及 SCA-1垣/ CD45垣/ CD31原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垣/ CD45垣/ CD31垣亚群中最高，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垣 CD45原 CD31原亚群中表达最高，而在 SCA-1垣 CD45垣 CD31垣亚群中表达很低，Itga4在未分选群中表达最高；Cxadr 在 SCA-1垣 CD45原 CD31原亚群中表达最高；Epor在 SCA-1垣 CD45原 CD31垣亚群中表达最高；myl3在 SCA-1垣 CD45垣 CD31垣亚群中表达最高。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。
목적：이용기인심편사선소서골수간충질간세포4개아군적차이표체기인，심조심기분화특이성기인。방법장소서골수간충질간세포통과 CD45화 CD31적자주분선，획득 SCA-1원/ CD45원/ CD31원、SCA-1원/ CD45원/ CD31원、SCA-1원/CD45원/ CD31원급 SCA-1원/ CD45원/ CD31원4군세포。채용기인심편기술완성 Agilent 소서전기인4*44K 심편표체보기인검측，분석차이표체기인。근거기인심편중소검측적기인표체량재 SCA-1원/ CD45원/ CD31원아군중최고，차위기타아군표체량적량배이상위원칙，통과 RT-PCR 험증각아군급미분선세포중적차이기인적표체량。결과근거피이삼관련산법계산4조세포아군지간적관련치위우0.979~0.995지간，통과 RT-PCR 검측발현，Rock2기인재 SCA-1원 CD45원 CD31원아군중표체최고，이재 SCA-1원 CD45원 CD31원아군중표체흔저，Itga4재미분선군중표체최고；Cxadr 재 SCA-1원 CD45원 CD31원아군중표체최고；Epor재 SCA-1원 CD45원 CD31원아군중표체최고；myl3재 SCA-1원 CD45원 CD31원아군중표체최고。결론 Rock2、Cxadr、Itga4、Epor 화myl3기인재간세포아군향심기분화중발휘불동작용。
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 miceˊwhole 4*44K 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.