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Expression of osterix inhibits bone morphogenetic protein-induced chondrogenic differentiation of mesenchymal progenitor cells

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Abstract

Osteoblasts and chondrocytes arise from common bipotential mesenchymal progenitor cells. Although the differentiation of these two cell lineages can be induced by treatment with bone morphogenetic proteins (BMPs), the responses of mesenchymal progenitors to BMP differ from cell line to cell line. Here we demonstrate that C3H/10T1/2 cells preferred chondrogenic differentiation, primary bone marrow stroma cells (MSCs) tended to convert to osteoblasts, and ST-2 cells differentiated into both the osteoblastic and chondrocytic lineages simultaneously, suggesting that a molecular switch functions to select cell fate. Osterix, the secondary master regulator of osteoblastogenesis, was induced by BMP at high and low levels in MSCs and ST-2 cells, respectively; in contrast, C3H/10T1/2 cells demonstrated only faint expression. As osterix has been suggested as a negative regulator of chondrogenesis, we hypothesized that the intense chondrocyte differentiation of C3H/10T1/2 cells may have resulted from an absence of osterix. We therefore restored osterix gene expression in C3H/10T1/2 cells using an adenovirus vector. Following BMP treatment, infection with an osterix-encoding virus dramatically inhibited the chondrocytic differentiation of C3H/10T1/2 cells, resulting instead in prominent osteoblast differentiation. These results indicate the chondrogenic potential of C3H/10T1/2 cells was abrogated by osterix expression. Chondrocyte differentiation of MSCs, however, was not enhanced by silencing the osterix gene using lentivirus-mediated shRNA, despite successful suppression of osteoblast differentiation. These results suggest that the low levels of osterix expression remaining after knockdown are sufficient to block chondrogenesis, whereas higher expression may be required to promote osteoblastic differentiation.

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Acknowledgments

This work was supported by grants-in-aid for scientific research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (T.I. and S.M.). We thank Y. Yuuki, A. Hanyu, N. Kaneniwa, and E. Kobayashi (The Cancer Institute) for their excellent technical assistance.

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Authors and Affiliations

  1. Department of Biochemistry, The Cancer Institute of the Japanese Foundation for Cancer Research, 3-10-6 Ariake, Koto-ku, Tokyo, 135-8550, Japan

    Hiroyuki Tominaga, Shingo Maeda & Takeshi Imamura

  2. Manipulation of Cell Fate, BioResource Center, RIKEN, Koyadai, Tsukuba, Japan

    Hiroyuki Miyoshi

  3. Department of Molecular Pathology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan

    Kohei Miyazono

  4. Department of Neuromusculoskeletal Disorder, Orthopaedic Surgery, Graduate School of Medicine and Dentistry, Kagoshima University, Kagoshima, Japan

    Hiroyuki Tominaga, Shingo Maeda & Setsuro Komiya

Authors
  1. Hiroyuki Tominaga
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  2. Shingo Maeda
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  3. Hiroyuki Miyoshi
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  4. Kohei Miyazono
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  5. Setsuro Komiya
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  6. Takeshi Imamura
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Corresponding author

Correspondence to Shingo Maeda.

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Tominaga, H., Maeda, S., Miyoshi, H. et al. Expression of osterix inhibits bone morphogenetic protein-induced chondrogenic differentiation of mesenchymal progenitor cells. J Bone Miner Metab 27, 36–45 (2009). https://doi.org/10.1007/s00774-008-0003-0

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  • DOI: https://doi.org/10.1007/s00774-008-0003-0

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