2005 Fiscal Year Final Research Report Summary
Strategic study on osteoblast and chondrocyte differentiation from embryonic stem (ES) cells
| Project/Area Number |
15390452
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | The University of Tokyo |
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Principal Investigator |
CHUNG Ung-il The University of Tokyo, Graduate School of Medicine, Associate Professor, 大学院・医学系研究科, 助教授 (30345053)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAGUCHI Hiroshi The University of Tokyo, Hospital, Associate Professor, 医学部附属病院, 助教授 (40282660)
NAKAMURA Kouzou The University of Tokyo, Hospital, Professor, 医学部附属病院, 教授 (60126133)
IKEDA Toshiyuki The University of Tokyo, Hospital, Visiting Assistant Professor, 医学部附属病院, 客員助手 (80322759)
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| Project Period (FY) |
2003 – 2005
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| Keywords | embryonic stem (ES) cells / osteoblast differentiation / chondrocyte differentiation / optimal differentiation condition / culture methods / regenerative medicine |
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| Research Abstract |
Mice expressing GFP transgenes under the control of promoter fragments of rat type I collagen gene and type U collagen gene were mated with wild-type mice, blastocytes were obtained and embryonic stem (ES) cells were isolated. After retroviruses expressing these transgenes were constructed, wild-type ES cells were infected with them and cell lines were established. ES cells from these mice, which fluoresce solely upon osteoblast or chondrocyte differentiation, were utilized as cell sensors for screening differentiation-inducing factors. We constructed adenoviruses expressing BMP receptors, Hh signaling molecule, Smo, Wnt transcription factor, LEF-1 and Runx2. Then, by exhaustively combining these osteoblast-differentiation inducing factors with above-mentioned ES cell sensor, we found that combination of Runx2 and BMP most strongly induced osteoblast differentiation.
As for cartilage-differentiation inducing factors, BMP and Sox signaling pathways were screened with the ES cell sensor a
… More
nd we found that combination of Sox9 and Sox5 or Sox6 most strongly induced cartilage differentiation.
Through these experiments, we optimized signaling for osteoblast or chondrocyte differentiation.
We also determined the optimal condition for proliferation of the ES cell lines in monolayer culture without differentiation (passaging intervals and best colony size). Aiming to use human ES cells in the near future, we purchased cynomolgus embryonic stem cells and gained experience in cultivating and differentiating them. We planted osteoblast-like and chondrocyte-like cells differentiated in the above determined optimal condition and cultured them on atelocollagen film. We succeeded in constructing sufficient matrix. We confirmed matrix calcification and metachromasie by toluidine blue staining. We transplanted these sheets to mouse bone and cartilage defects. One-four months after transplantation, we euthanized them after anesthesia and made histological and immunohistological examinations of the transplanted parts, through which we observed good regeneration in the bone-defect model but no significant hearing effects in the cartilage-defect model. We speculate that cartilage differentiation signal is not fully optimized. We plan to continue screening, considering various combinations with other signaling pathways. Less
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[Journal Article] Osteoarthritis development in novel exper imental mouse models induced by knee joint instability.2005
Author(s)
Kamekura S, Hoshi K, Shimoaka T, Chung U, Chikuda H, Yamada T, Uchida M, Ogata N, Seichi A, Nakamura K, Kawaguchi H.
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Journal Title
Osteoarthritis Cartilage 13(7)
Pages: 632-641
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] The canonical Wnt signaling pathway promotes chondrocyte differentiation in a Sox9-dependent manner.2005
Author(s)
Yano F, Kugimiya F, Ohba S, Ikeda T, Chikuda H, Ogasawara T, Ogata N, Takato T, Nakamura K, Kawaguchi H, Chung U.
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Journal Title
Biochem Biophys Res Commun 333(4)
Pages: 1300-1308
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Involvement of endogenous bone morphogenetic protein (BMP)2 and BMP6 in bone formation.2005
Author(s)
Kugimiya F, Kawaguchi H, Kamekura S, Chikuda H, Ohba S, Yano F, Ogata N, Katagiri T, Harada Y, Azuma Y, Nakamura K, Chung U.
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Journal Title
J Biol Chem 280
Pages: 35704-35712
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Regenerative Therapy of Bone / Cartilage2005
Author(s)
Oba Shinsuke, Igawa Kazuyo, Nakamura Kozo, Kawaguchi Hiroshi, Takatou Takeshi, Chung, Ung-il
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Journal Title
Geriatrics Update 2005-2006 (Edited by Japan Geriatrics Society Journal Editing Committee) (Medicalview company)
Pages: 199-206
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Distinct Role of Smad Pathways and p38 Pathways in Cartilage-specific Gene Expression in Synovial Fibroblasts.2004
Author(s)
Seto H, Kamekura S, Miura T, Yamamoto A, Chikuda H, Ogata T, Hiraoka H, Nakamura K, Kurosawa H, Chung U, Kawaguchi H, Tanaka S.
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Journal Title
J Clin Invest 113
Pages: 718-726
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] PPARg insufficiency enhances osteogenesis from bone marrow progenitors.2004
Author(s)
Akune T, Oba S, Kamekura S, Yamaguchi M, Chung U, Kubota N, Terauchi Y, Harada Y, Azuma Y, Kadowaki T, Nakamura K, Kawaguchi H.
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Journal Title
J Clin Invest 113
Pages: 846-855
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] SRC-1 is necessary for skeletal responses to sex hormones in both males and females.2004
Author(s)
Yamada T, Kawano H, Sekine K, Matsumoto T, Fukuda T, Azuma Y, Chung U, Chambon P, Nakamura K, Kato S, Kawaguchi H.
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Journal Title
J Bone Miner Res 19
Pages: 1452-1461
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] The combination of SOX5, SOX6 and SOX9 (the SOX trio) provides signals sufficient for induction of permanent cartilage.2004
Author(s)
Ikeda T, Kawano H, Sekine K, Matsumoto T, Fukuda T, Azuma Y, Chung U, Chambon P, Nakamura K, Kato S, Kawaguchi H.
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Journal Title
Arthritis Rheum 50
Pages: 3561-3573
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Cyclic GMP-dependent kinase II is a molecular switch from proliferation to hypertrophic differentiation of chondrocytes.2004
Author(s)
Chikuda H, Kugimiya F, Ikeda T, Ogasawara T, Shimoaka T, Kawano H, Kamekura S, Tsuchida A, Yokoi N, Nakamura K, Komeda K, Chung U, Kawaguchi H.
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Journal Title
Genes Dev 18
Pages: 2418-2429
Description
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