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Establishment of Assay System for Studying Osteoblastic Function and Differentiation Using Newly Isolated Osteoblast-Like Cell Lines

Research Project

Project/Area Number 01870076
Research Category

Grant-in-Aid for Developmental Scientific Research (B).

Allocation TypeSingle-year Grants
Research Field Morphological basic dentistry
Research InstitutionShowa University

Principal Investigator

YOSHIKI Shusaku  School of Dentistry, Professor Showa University, 歯学部, 教授 (30085740)

Co-Investigator(Kenkyū-buntansha) IKEDA Tohru  School of Dentistry Showa University Research Associate, 歯学部, 助手 (00211029)
TAKAHASHI Naoyuki  School of Dentistry Showa University Assistant Professor, 歯学部, 講師 (90119222)
SASAKI Takahisa  School of Dentistry, Showa University Associate Professor, 歯学部, 助教授 (50129839)
KAKUTA Saburou  School of Dentistry, Showa University Associate Professor, 歯学部, 助教授 (40112726)
YAMAGUCHI Akira  School of Dentistry, Showa University Associate Professor, 歯学部, 助教授 (00142430)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥7,300,000 (Direct Cost: ¥7,300,000)
Fiscal Year 1990: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1989: ¥4,400,000 (Direct Cost: ¥4,400,000)
Keywordsosteoblast / differentiation / cell line / BMP / retinoic acid / 骨肉腫
Research Abstract

In order to establish the suitable assay system for studying osteoblastic phenotypes and differentiation, we isolated various kinds of clonal osteoblast-like cell lines from neonatal rat calvariae and human osteosarcoma.
I. Establishment of osteoblast-like cell lines
1) Rat osteoblast-like cell lines isolated from rat calvariae
Five clonal osteoblast-like cell lines (ROB-C8a, Cll, C20. C23. C26) were isolated from 1-day old rat calvariae. Cll, C20 and C23 cell lines expressed high degree of osteoblastic properties but C26 cells expressed very low level of that properties. Expression degree of the osteoblastic properties in C8a was intermediate. More importantly, C26 cells expressed differentiation potential into not only osteoblasts but also myotubes and adipocytes. These indicated that C26 cells are useful cell line for studying the differentiation pathway of osteoblasts, muscle cells and adipocytes from undifferentiated mesenchymal cells.
2) Human osteoblast-like cell lines isolated from … More human osteosarcoma
Two clonal osteoblast-like cell lines (YR-2 and YR-6) were isolated from human osteosarcoma. Osteoblastic expression of these cell lines were compared with those of other established osteosarcoma cell lines (HOS, SAOS-2 and Mg63). This study demonstrated the diversity of osteoblastic phenotypes among various human osteosarcoma cell lines tested, and suggested that much understanding of the osteoblastic characteristics in each cell line is important when human osteosarcoma cell lines are used for studying osteoblastic phenotypes.
II. Regulatory mechanism of osteoblast differentiation
We demonstrated that bone morphogenetic protein (BMP) and retinoic acid play an important role in osteoblast differentiation.
1) BMP : Recombinant human bone morphogenetic protein-2 (rhBMP-2 ; provided from Dr. Wozney, Genetics Institute, MA, USA) strongly stimulated osteoblastic differentiation and inhibited myogenic differentiation in the immature osteoblast-like cell line, C26. rhBMP-2 also induced non-osteogenic fibroblasts, C3H1OT1/2 cells, to differentiate into osteoblastic cells. These results strongly suggest that BMP-2 plays an important role in osteoblastic differentiation from undifferentiated mesenchymal cells.
2) Retinoic acid : Retinoic acid stimulated ALP activity and PTH responsiveness, and inhibited myogenic and adipose differentiation in C26 cells. These results suggest that retinoic acid regulates differentiation pathway into osteoblasts, muscle cells and adipocytes from undifferentiated mesenchymal cells. Less

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (31 results)

All Other

All Publications (31 results)

  • [Publications] 山口 朗: "骨芽細胞の培養." 組織培養.15. 155-159 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 山口 朗: "培養骨芽細胞の多様性." 日本骨代謝学会雑誌.7. 16-20 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 山口 朗: "骨芽細胞の分化と機能." ホルモンと臨床. 38. 15-19 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Katagiri,K: "The nonーosteogenic mouse pluripotential cell line, C3H10T1/2, is induced to differentiate into osteoblastic cells by recombinant human bone morphogenetic proteinー2." Biochem.Biophys.Res,Commun.172. 295-299 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishimi,Y.: "lLー6 is produced by osteoblasts and induces bone resorption." J.Immunol.145. 3297-3303 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 片桐 岳信: "骨芽細胞の分化." 最新医学. 46. 206-212 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Akira Yamaguchi, Tohru Ikeda, Shusaku Yoshiki: "Culture of osteoblasts (in Japanese)" Soshikibaiyou. 15. 155-159 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Akira Yamaguchi: "Diversity of osteoblast-like cells in vitro. (in Japanese)." Jap. J. Bone Miner. Res.7. 16-20 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Udagawa, N., Takahashi, N., Akatsu, T., Sasaki, T., Yamaguchi, A., Kodama, H., Martin, T. J., and Suda, T.: "The bone marrow-derived stromal cell lines MC3T3-G2/PA6 and ST2 support osteoblast-like cell differentiation on cocultures with nouse spleen cells." Endocrinology. 125. 1805-1813 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Katagiri, K., Yamaguchi, A., Ikeda, T., Yoshiki, S., Wozney, J. M., Rosen, V., Wang, E. A., Tanaka, H., Omura, S., and Suda, T.: "The non-osteogenic mouse pluripotential cell line, C3H10T 1/2, is induced to differentiate into osteoblastic cells by recombinant human bone morphogenetic protein-2." Biochem. Biophys. Res. Commun.172. 295-299 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nojima, N., Kobayashi, M., Shionome, M., Takahashi, N., Suda, T., and Hasegawa, T.: "Fibroblastic cells derived from bovine periodontal ligaments have the phenotypes of osteoblasts." J. Periodont. Res.25. 179-185 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ozawa, H., Imamura, K., Abe, E., Takahashi, N., Hiraide, T., Shibasaki, Y., Fukuhara, T. and Suda, T.: "Effect of a continuopusly applied compressive pressure on mouse osteoblast-like cells (MC3T3-E1) in vitro." J. Cell Physiol.142. 177-185 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Imamura, K., Ozawa, H., Hiraide, T., Takahashi, N., Shibasaki, Y., Fukuhara, T., and Suda, T.: "Continuously applied compressive pressure induces bone resorption by a mechanism involving prostaglandin E2 synthesis." J. Cell Physiol.144. 222-228 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Shinki, T., Tanaka, H., Takito, J., Yamaguchi, A., Nakamura, Y., Yoshiki, S., and Suda, T.: "Putrescine is involved in the vitamin D action in chick intestine." Gastroenterology. 100. 113-122 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishimi, Y., Miyaura, C., Jin, C. H., Akatsu, T., Abe, E., Nakamura, Y., Yamaguchi, A., Yoshiki, S., Matsuda, T., Hirano, T., Kishimoto, T., and Suda, T.: "IL-6 is produced by osteoblasts and induces bone resorption." J. Immunol.145. 3297-3303 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Yamaguchi, A., and Kahn, A. J.: "Clonal osteogenic cell lines express myogenic and adipocytic developmental potential." Calcif. Tissue Int. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Akatsu, T., Takahashi, N., Udagawa, N,. Imamura, K., Yamaguchi, A., Sato, K., Nagata, N., and Suda, T.: "The role of prostaglandins in interleukin 1 (IL-1)ーinduced bone resorption in mice." J. Bone Miner. Res. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Yamaguchi, A., Katagiri, T., Ikeda, T., Wozney, J. M., Rosen, V., Wang, E, A., Kahn, A, J., Suda, T., and Yoshiki, S.: "Recombinant human bone morphogenetic protein-2 stimulates osteoblastic maturation and inhibits myogenic differentiation in vitro." J. Cell Biol.(1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nakamura. Y., Yamaguchi, A., Ikeda, T., and Yoshiki, S.: "Acid phosphatase activity is detected exclusively in the osteoblastic lineage by pre-treatment with cyanuric chloride." J. Histochem. Cytochem.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 山口 朗: "骨芽細胞の培養." 組織培養.15. 155-159 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] 山口 朗: "培養骨芽細胞の多様性." 日本骨代謝学会雑誌.7. 16-20 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] 山口 朗: "骨芽細胞の分化と機能." ホルモンと臨床. 38. 15-19 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Katagiri,K: "The nonーosteogenic mouse Pluripotential cell line,C3H10T1/2,is induced to differentiate into osteoblastic cells by recombinant human bone morphogenetic proteinー2." Biochem.Biophys.Res.Commun.172. 295-299 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Ishimi,Y.: "ILー6 is produced by osteoblasts and induces bone resorption." J.Immunol.145. 3297-3303 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 片桐 岳信: "骨芽細胞の分化." 最新医学. 46. 206-212 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Udagawa,N.,Takahashi,N.,Akatsu T.,Sasaki,T.,Yamaguchi,A.,Kodama,H.,Martin,T.J.,Suda,T.: "The bone marrow-derived stromal cell lines MC3T3-G2/PA6 and ST2 support osteoclastlike cell differentiation in cocultures with mouse spleen cells." Endocrinology. 125. 1805-1813

    • Related Report
      1989 Annual Research Report
  • [Publications] 吉木周作: "脱灰切片による硬組織内物質の組織形態的観察法に関する一連の研究." 昭和歯学会雑誌. 8. 247-255 (1988)

    • Related Report
      1989 Annual Research Report
  • [Publications] 山口朗,吉木周作: "骨芽細胞の形態と機能の多様性." 臨床科学. 25. 241-248 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 山口朗,吉木周作: "骨のコラゲナ-ゼ." 骨代謝誌. 7. 50-57 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 山口朗,池田通,吉木周作: "骨芽細胞の培養." 組織培養. 15. 155-159 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Yamaguchi,A.,Kahn,A.J.: "Clonal osteogenic cell lines express myogenic and adipocytic developmental potential."

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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