体外培养大鼠骨髓间充质干细胞成骨基因表达谱的检测

目的:探讨体外培养大鼠骨髓间充质干细胞(MSC)成骨相关基因的表达谱,明确体外培养大鼠骨髓MSC的成骨潜能。方法:体外培养SD大鼠骨髓MSC,取第3代细胞提取总核糖核酸(RNA),制备杂交探针,并运用SuperArray公司提供的大鼠Oligo GEArray骨再生基因芯片(每张芯片可测113种基因)进行杂交,化学发光检测,通过X线胶片或台式扫描获得化学发光的芯片图像,使用网上提供的综合型GEArry表达分析配套软件(GEArryExpression Analysis Suite)进行完整的芯片数据分析。结果:检测体外培养大鼠骨髓MSC的113种成骨相关的基因,其中有31种基因表达强度较低,45种基因呈中等强度表达,37种基因表达强度较高。结论:体外培养大鼠骨髓MSC具有部分成骨细胞基因表型特征,但成熟成骨细胞的特征基因表达量较低,提示大鼠MSC是具有较强成骨细胞分化潜能的成纤维样细胞。

Detection of the osteogenesis gene expression of rat bone marrow mesenchymal stem cells using gene chip

Objective:To investigate the osteogenesis genes expressed in rat bone marrow mesenchymal stem cells(MSC) and identify their potential of osteogenesis.Methods:SD rat bone marrow MSCs were isolated and cultured in vitro and total RNA extracted and purified from the third-passage cells.The probe was synthesized and the gene profile of the MSCs examined by using the rat oligo osteogenesis microarray,which contained 113 genes involved in the regulation of osteogenic differentiation.The hybridization signals were acquired using X-ray film by chemiluminescent detection and the data obtained were analyzed through the web-based completely integrated GEArray Expression Analysis Suite.Results:Among the 113 osteogenesis genes,31 expressed lowly,45 moderately and 37 highly.Conclusion:Some osteoblast marker genes were expressed in the rat bone marrow MSCs,but with a low expression of the mature osteoblast marker genes,which indicates that rat bone marrow MSCs have the potential of osteoblast differentiation.