Identification Of Genes Differently Expressed In Human Bone Marrow Mesenchymal Stem Cells Between Early And Late Passages

Mesenchymal stem cells (MSCs) can self-renew, are multipotent cells and can differentiate into many different cells. They are important cell souece for tissue engineering and cell therapy. So far, little is known about how MSC growth and differentiation is regulated, and this study tried to gain some insight into this question by identification of genes differentially expressed in human bone marrow mesenchymal stem cells (hBM-MSCs) between early passages and late passages. hBM-MSCs at passage 5 and passage 20 were used as a tester and a driver respetively for subtractive hybridization experiment. PCR product derived from subtractive hybridization was cloned into pGEM-Teasy vector. A total of 117 positive clones were taken for DNA sequencing. DNA sequence of each clone was compared with GenBank database for gene homologes, and 21 different human known genes were identified representing the 117 clones. DNA sequence homologies between each of these 21 genes and their respective known genes were greater than 90%. To confirm the differential expression of these genes, semi-quantitative RT-PCR analysis were performed, and it showed that 6 genes were indeed differentially expressed in hBM-MSCs between early passages and late passages by more than 1 fold. These genes included rab23, ddx21, tnfaip3, nomo2, pabpc3 and cnn3. These genes may play an important role in maitaning and regulating hBM-MSCs growth and differentiation, and may provide useful information for controlling hBM-MSCs growth and differentiation for tissue engineering.