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Comparative sequential morphological analyses during in vitro chondrogenesis and osteogenesis of mesenchymal stem cells embedded in collagen gels

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Abstract

Chondrogenesis and osteogenesis during fetal development and postnatal growth constitute one of the most interesting and complicated subjects in biology. In this study, bone marrow mesenchymal stem cells (MSCs) were embedded in collagen gel, cultured in chondrogenic or osteogenic medium, and compared morphologically during chondrogenic and osteogenic differentiation sequentially by light and electron microscopy and immunohistochemical examination. Before induction, the MSCs were dispersed and round in the collagen gel. At day 1, MSCs with a large number of short processes produced extracellular fibers whose immunoreactivity was positive for collagen type I. At day 3, the shape of MSCs changed from round to elongated. Gap junctions positive for connexin 43 were also observed. At day 7, remarkable morphological differences were first observed during chondrogenesis and osteogenesis. The shape of MSCs changed to polygonal without cell processes during chondrogenesis, while MSCs remained spindle shaped with long processes during osteogenesis. Concurrently, collagen type II during chondrogenesis and osteocalcin during osteogenesis were first detected. At day 21, chondrogenesis and osteogenesis of the MSC/collagen composite further progressed, respectively. In vitro chondrogenesis and osteogenesis using an MSC/collagen composite clarified the morphological differences.

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Acknowledgments

We thank Kenjiro Wake and Hisae Hori for valuable discussions, Akiko Yokoyama for help with in vitro chondrogenesis of MSCs, and Yuko Kawamura for preparing the figures. This study was supported by grants from the Japan Society for the Promotion of Science (19500403) to Shizuko Ichinose, from the Japanese Ministry of Education Global Center of Excellence (GCOE) Program, International Research Center for Molecular Science in Tooth and Bone Diseases to Takeshi Muneta, and from the Health and Labor Sciences Research Grant, Research on Regenerative Medicine for Clinical Application to Ichiro Sekiya.

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No competing financial interests exist.

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Authors and Affiliations

  1. Research Center for Medical and Dental Sciences, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan

    Shizuko Ichinose

  2. Department of Medicine, Sanraku Hospital, Tokyo, Japan

    Motoki Tagami

  3. Section of Orthopedic Surgery, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan

    Takeshi Muneta

  4. Global Center of Excellence Program for International Research Center for Molecular Science in Tooth and Bone Disease, Tokyo Medical and Dental University, Tokyo, Japan

    Takeshi Muneta

  5. Department of Oral Surgery, Minato Red Cross Hospital, Yokohama, Japan

    Hitoshi Mukohyama

  6. Section of Cartilage Regeneration, Graduate School, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan

    Ichiro Sekiya

Authors
  1. Shizuko Ichinose
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  2. Motoki Tagami
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  3. Takeshi Muneta
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  4. Hitoshi Mukohyama
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  5. Ichiro Sekiya
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Correspondence to Shizuko Ichinose or Ichiro Sekiya.

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Ichinose, S., Tagami, M., Muneta, T. et al. Comparative sequential morphological analyses during in vitro chondrogenesis and osteogenesis of mesenchymal stem cells embedded in collagen gels. Med Mol Morphol 46, 24–33 (2013). https://doi.org/10.1007/s00795-012-0005-9

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  • DOI: https://doi.org/10.1007/s00795-012-0005-9

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