Analysis of stem-cell differentiation mechanism by using DNA microarray and novel bisulfite modification.

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We performed DNA microarray analysis of immortalized human mesenchymal stem cell clones. Among them, clone no.12 showed highest ability of differentiation to osteoblasts, chondrocytes and adipocytes, and its profiling analysis showed characteristic patterns of gene expression. In order to know the relationship between stem cell differentiation and DNA methylation, we next performed global methylation analysis of stem cell clones under differentiation processes by using CpG methylation array system. As a result, we isolated 11 genes under osteoblastic differentiation process and four genes under adipocytic differentiation process as candidates of up-regulated or down-regulated genes of methylation status. Furthermore, we isolated 82 genes varying their methylation levels during stem cell differentiation by using novel bisulfite modification with which genome-wide identification of methylation sites may become possible. These genes were supposed to be important for regulation of stem cell differentiation and reconstruction of methylation status. These results may contribute to the progress of stem cell research and regeneration medicine up to date.