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Differentiation of Human Bone Marrow Mesenchymal Stem Cells to Chondrocytes for Construction of Three-dimensional Cartilage Tissue

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Abstract

A differentiation method of human bone marrow mesenchymal stem cells (MSCs) to chondrocytes was developed for the construction of a three-dimensional (3D) cartilage tissue. The adhesive cells, which were isolated from a human bone marrow aspirate were embedded in type I collagen in a poly-l-lactate-glycolic acid copolymer (PLGA) mesh and cultivated for 4 week together with growth factors. The degree of cellular differentiation was estimated by quantitative RT-PCR of aggrecan and type II collagen mRNAs and by staining with Safranin O. The 3D culture showed a higher degree of differentiation even without growth factors than the conventional pellet culture with growth factors, namely, dexamethasone and transforming growth factor (TGF)-β 3. The 3D culture for 2 week with the combined addition of dexamethasone, TGF-β 3, and insulin-like growth factor (IGF)-I reached a 30% expression of aggrecan mRNA compared with that in primary human chondrocytes, while the aggrecan mRNA expression in the conventional pellet culture was less than 2%. The sequential two-step differentiation cultivation, during which the cells were cultivated in 3D for 1 week after the conventional two-dimensional (2D) culture for 1 week, could markedly accelerate the expression of aggrecan mRNA compared with the 3D cultivation for 2 week.

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Authors and Affiliations

  1. International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita, Osaka, 565-0871, Japan

    Chikayoshi Matsuda, Mutsumi Takagi & Toshiomi Yoshida

  2. Department of Orthopedic Surgery, Osaka-Minami National Hospital, 2-1 Kidohigashi-machi, Kawachinagano, Osaka, 586-8521, Japan

    Takako Hattori

  3. Orthopedic Surgery, Shinshu University School of Medicine, 3-1-1 Asahi, Matsumoto, Nagano, 390-8621, Japan

    Shigeyuki Wakitani

  4. Division of Biotechnology and Molecular Chemistry, Graduate School of Engineering, Hokkaido University, Sapporo, 060-8628, Japan

    Mutsumi Takagi

Authors
  1. Chikayoshi Matsuda
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  2. Mutsumi Takagi
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  3. Takako Hattori
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  4. Shigeyuki Wakitani
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  5. Toshiomi Yoshida
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Correspondence to Mutsumi Takagi.

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Matsuda, C., Takagi, M., Hattori, T. et al. Differentiation of Human Bone Marrow Mesenchymal Stem Cells to Chondrocytes for Construction of Three-dimensional Cartilage Tissue. Cytotechnology 47, 11–17 (2005). https://doi.org/10.1007/s10616-005-3751-x

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  • DOI: https://doi.org/10.1007/s10616-005-3751-x

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