Gene Expression Analyses of Human Mesenchymal Stem Cells Cultured in Osteogenic Differentiation Medium for 3, 7, 14 and 21 Days by Genome Focus DNA Microarray and Real-time PCR

Abstract

The purpose of this study was to evaluate gene expressions of human mesenchymal stem cells (hMSC) cultured in osteogenic differentiation medium (OM) which contained ascorbic acid, β-glycerophosphate and dexamethasone for 0 (control), 3, 5, 7, 14 and 21 days by 8.5k Genome Focus DNA microarray and real-time PCR. It was confirmed by the DNA microarray analysis that 327 genes of hMSC were significantly up-regulated by culture in OM especially at 14 and 21 days while 156 genes were down-regulated. Up-regulated genes included osteoblast-related genes such as secreted phosphoprotein 1 (osteopontin) gene and hypothetical protein expressed in osteoblast gene, along with angiogenesis-related genes and cell cycle arrestrelated genes, while down-regulated genes contained stroma-related genes and keratin-related genes. Expressions of several osteogenic differentiation marker genes such as (down-regulated) osteonectin gene and (up-regulated) BMP2 gene were also evaluated by real-time PCR. Gene expression database identified here might contribute to dental tissue engineering.