论文部分内容阅读
目的:探讨小鼠间充质干细胞(MSCs)定向诱导分化成脂肪细胞微小RNA(miRNA)表达的变化,为进一步研究miRNA调控MSCs向脂肪细胞分化的分子机制奠定基础。方法:采用全骨髓体外分离结合差速贴壁法纯化扩增C57BL/6小鼠MSCs,形态学观察细胞生长情况,并用免疫组化方法鉴定细胞表面抗原CD29、CD44和CD34的表达。脂肪细胞分化诱导剂诱导MSCs分化为脂肪细胞,利用油红O染色,判断MSCs成脂分化情况。运用miRNA芯片技术检测MSCs和脂肪细胞中差异表达的miRNA。结果:①倒置显微镜下观察,传5代后可获得均一性较高的MSCs;免疫组化显示90%以上的骨髓间质干细胞CD29、CD44阳性,CD34阴性。MSCs经脂肪诱导剂诱导后,胞内大量脂滴形成,油红O染色阳性;②基因微阵列分析表明,小鼠MSCs分化成脂肪细胞差异表达的miRNA共75个,其中20个表达上调、55个表达下调。结论:MSCs分化成脂肪细胞存在miRNA表达的变化,某些miRNA很可能具有重要的调控MSCs成脂分化的作用。
OBJECTIVE: To investigate the changes of miRNA expression in mouse adipose-derived mesenchymal stem cells (MSCs) induced by differentiation-induced differentiation, which lays the foundation for further study on the molecular mechanism of miRNA regulation of MSCs differentiation into adipocytes. Methods: MSCs of C57BL / 6 mice were purified by whole bone marrow in vitro combined with differential adherent method. Morphology was used to observe the cell growth. The expression of CD29, CD44 and CD34 were identified by immunohistochemistry. Adipocyte differentiation inducer induced MSCs to differentiate into adipocytes. Oil red O staining was used to determine the adipogenic differentiation of MSCs. Using miRNA microarray technique to detect differentially expressed miRNAs in MSCs and adipocytes. Results: ① Under inverted microscope, MSCs with higher homogeneity were obtained after passage 5 for 5 generations. Immunohistochemistry showed that more than 90% of MSCs were positive for CD29, CD44 and negative for CD34. MSCs were induced by lipid inducer, a large number of intracellular lipid droplets formed, oil red O staining; ② gene microarray analysis showed that MSCs differentiate into adipocytes differentially expressed miRNA a total of 75, of which 20 were up-regulated, 55 A decrease in expression. Conclusion: The differentiation of MSCs into adipocytes has the miRNA expression changes. Some miRNAs may play an important role in regulating adipogenic differentiation of MSCs.