Journal of Oral Tissue Engineering

ORIGINAL ARTICLE
 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

Masayuki TAIRA1, Naoyuki CHOSA1, Kaori SASAKI1, Setsuo SAITOH1, Takashi NEZU1, Nobuko SATO2 and Yoshima ARAKI1

1Department of Dental Materials Science and Technology, 2Department of Biochemistry, Iwate Medical University School of Dentistry, Iwate, Japan.

J Oral Tissue Engin 2007;5(1): 35-47
Full Text. DOI https://doi.org/10.11223/jarde.5.35
SYNOPSIS
The purpose of this study was to evaluate gene expressions of human mesen-chymal 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.

Key words: Mesenchymal stem cells, Osteogenic differentiation medium, Gene expression, DNA microarray, Real-time PCR