Isolation and characterization of bone marrow-derived mesenchymal progenitor cells with myogenic and neuronal properties

Mitsutaka Shiota, Toshio Heike, Munetada Haruyama, Shiro Baba, Atsunori Tsuchiya, Hisanori Fujino, Hirohiko Kobayashi, Takeo Kato, Katsutsugu Umeda, Momoko Yoshimoto, Tatsutoshi Nakahata

Research output: Contribution to journalArticlepeer-review

73 Scopus citations


Sphere formation has been utilized as a way to isolate multipotent stem/progenitor cells from various tissues. However, very few studies on bone marrow-derived spheres have been published and assessed their multipotentiality. In this study, multipotent marrow cell populations were isolated using a three-step method. First, after elimination of hematopoietic cells, murine marrow-derived adherent cells were cultured in plastic dishes until small cells gradually appeared and multiplied. Cells were then cultured under non-adherent conditions and formed spheres that were immunopositive for a neural precursor marker, nestin. RT-PCR analysis also revealed that the spheres were positive for nestin in addition to PPARγ, osf2, SOX9, and myoD, which are markers of precursors of adipocytic, osteoblastic, chondrocytic, and skeletal myeloblastic lineages, respectively. Finally, spheres were dissociated into single cells and expanded in adherent cultures. Under appropriate induction conditions, the sphere-derived cells acquired the phenotypic properties in vitro of neurons, skeletal myoblasts, and beating cardiomyocytes, as well as adipocytes, osteoblasts, and chondrocytes. Next, sphere-derived cells were transplanted into murine myocardial infarction models. One month later, they had become engrafted as cardiomyocytes, and cardiac catheterization showed significant functional improvements. Thus, sphere-derived cells represent a new approach to enhance the multi-differentiation potential of murine bone marrow.

Original languageEnglish (US)
Pages (from-to)1008-1023
Number of pages16
JournalExperimental Cell Research
Issue number5
StatePublished - Mar 10 2007


  • Acute myocardial infarction
  • Cardiomyocytes
  • Mesenchymal stem cells
  • Multipotent spheres
  • Murine bone marrow
  • Neurons
  • Skeletal myoblasts

ASJC Scopus subject areas

  • Cell Biology

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