[Screening differentially expressed genes in human bone marrow stromal cells at defined stage of differentiation.].

To screen differentially expressed genes involved in osteogenic differentiation of human bone marrow stromal cells (BMSCs) at defined stages, subtractive cDNA library was established by means of suppression subtractive hybridization. The BMSCs cultured for 12 and 21 d were used as driver and tester, respectively. A subtract library was successfully constructed and five positive clones were selected from the library. Sequencing analysis and homology comparison showed that the five clones differentially expressed in BMSCs cultured for 21 d were at least 90% homologous with the known genes in human GenBank. It was interestingly found that the osteogenic BMSCs cultured for 21 d differentially expressed decorin and Bax inhibitor 1. RT-PCR was performed to confirm the differentially expressed genes. The results showed that the expression of Bax inhibitor 1 was significantly higher in the cells of 21-day than that of 12-day, while the expression of decorin was only detected in the cells of 21-day.

为了在定义的阶段筛选参与人骨髓基质细胞（BMSC）成骨分化的差异表达基因，通过抑制减法杂交建立了减法cDNA文库。培养12和21 d的BMSC分别用作驱动器和测试仪。成功地构建了减去库，并从图书馆中选择了五个阳性克隆。测序分析和同源性比较表明，在培养的BMSC中差异表达21 d的五个克隆与人类GenBank中已知基因至少同源90％。有趣的是，对21 d差异表达的装饰蛋白和BAX抑制剂1培养的成骨BMSC进行了。进行RT-PCR以确认差异表达的基因。结果表明，在21天的细胞中，BAX抑制剂1的表达显着高于12天的表达，而Decorin的表达仅在21天的细胞中被检测到。