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

2006 Fiscal Year Final Research Report Summary

• Project/Area Number: 17590080
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Shujitsu University
• Principal Investigator: NAKANISHI Tohru Shujitsu University, School of Pharmacy, Professor, 薬学部, 教授 (30243463)
• Co-Investigator(Kenkyū-buntansha): HAYATSU Hikoya Shujitsu University, School of Pharmacy, Professor, 薬学部, 教授 (10012593) ; MORI Hiroki Shujitsu University, School of Pharmacy, Assistant, 薬学部, 助手 (40388989) ; OKA Tsuyoshi Okayama Univ., Graduate School of Medicine, Dentistry and Pharmaceutical Science, Assistant, 大学院医歯薬学総合研究科, 助手 (50160651)
• Project Period (FY): 2005 – 2006
• Keywords: stem cells / telomerase / DNA chips / DNA methylation / osteoblasts / adipocytes

Research Abstract

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.

Research Products

• [Journal Article] Characterization and gene expression profiling of human mesenchymal stem cells. (2007)
  • Author(s): Mori H et al.
  • Journal Title: J Hard Tissue Biol 16(In press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Characterization and gene expression profiling of human mesenchymal stem cells. (2007)
  • Author(s): Mori H, Itano Y, Kobayashi N, Kosaka Y, Tanaka N, Nakanishi T.
  • Journal Title: J Hard Tissue Biol 16 (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Expression of WISP-1(ccn4), WISP-2(ccn5) and WISP-3(ccn6) in rheumatoid arthritic synovium evaluated by DNA microarrays. (2006)
  • Author(s): Mori H et al.
  • Journal Title: J Hard Tissue Biol 15 Pages: 89-95
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Bisulfite modification of cytosine and 5-methylcytosine as used in epigenetic studies. (2006)
  • Author(s): Hayatsu H
  • Journal Title: Genes and Envir 28 Pages: 1-8
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Expression of WISP-1(ccn4), WISP-2(ccn5) and WISP-3(ccn6) in rheumatoid arthritic synovium evaluated by DNA microarrays. (2006)
  • Author(s): Mori H.Nishida K, Ozaki T, Inoue H, Setsu K, Tsujigiwa H, Nagatsuka H, Gunduz M, Nakanishi T.
  • Journal Title: J Hard Tissue Biol 15 Pages: 89-95
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Bisulfite modification of cytosine and 5-methylcytosine as used in epigenetic studies. (2006)
  • Author(s): Hayatsu H.
  • Journal Title: Genes and Envir 28 Pages: 1-8
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Induction of claudin-4 by nonsteroidal anti-inflammatory drugs and its contribution to their chemopreventive effect. (2005)
  • Author(s): Mima S et al.
  • Journal Title: Cancer Res 65 Pages: 1868-1876
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Gene expression of connective tissue growth factor (CTGF/CCN2) in calcifying tissues of normal and cbfa1-null mutant in late stage of mouse embryonic development. (2005)
  • Author(s): Yamaai T et al.
  • Journal Title: J Bone Miner Metab 23 Pages: 280-288
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Gene expression profiling and differentiation of immortalized human mesenchymal stem cells. (2005)
  • Author(s): Mori H et al.
  • Journal Title: J Hard Tissue Biol 14 Pages: 267-268
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Induction of claudin-4 by nonsteroidal anti-inflammatory drugs and its contribution to their chemopreventive effect. (2005)
  • Author(s): Mima S, Tsutsumi S, Ushijima H, Takeda M, Fukuda I, Yokomizo K, Suzuki K, Sano K, Nakanishi T, Tomisato W, Tsuchiya T, Mizushima T.
  • Journal Title: Cancer Res 65 Pages: 1868-1876
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Gene expression of connective tissue growth factor (CTGF/CCN2) in calcifying tissues of normal and cbfal-null mutant in late stage of mouse embryonic development. (2005)
  • Author(s): Yamaai T, Nakanishi T, Asano M, Nawachi K, Yoshimichi G, Ohyama K, Komori T, Sugimoto T, Takigawa M.
  • Journal Title: J Bone Miner Metab 23 Pages: 280-288
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Gene expression profiling and differentiation of immortalized human mesenchymal stem cells. (2005)
  • Author(s): Mori H, Kato J, Matsuo H, Kobayashi N, Kosaka Y, Oka T, Tanaka N, Nakanishi T.
  • Journal Title: J Hard Tissue Biol 14 Pages: 267-268
  • Description: 「研究成果報告書概要(欧文)」より