title: Isolation and characterization of bone marrow-derived mesenchymal progenitor cells with myogenic and neuronal properties.
abstract: 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 PPARgamma, 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.
authors.affiliation: Department of Pediatrics, Graduate School of Medicine, Kyoto University, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Mitsutaka
authors.initials: M
authors.lastname: Shiota
authors.forename: Toshio
authors.initials: T
authors.lastname: Heike
authors.forename: Munetada
authors.initials: M
authors.lastname: Haruyama
authors.forename: Shiro
authors.initials: S
authors.lastname: Baba
authors.forename: Atsunori
authors.initials: A
authors.lastname: Tsuchiya
authors.forename: Hisanori
authors.initials: H
authors.lastname: Fujino
authors.forename: Hirohiko
authors.initials: H
authors.lastname: Kobayashi
authors.forename: Takeo
authors.initials: T
authors.lastname: Kato
authors.forename: Katsutsugu
authors.initials: K
authors.lastname: Umeda
authors.forename: Momoko
authors.initials: M
authors.lastname: Yoshimoto
authors.forename: Tatsutoshi
authors.initials: T
authors.lastname: Nakahata
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001854:Bone Marrow Cells / Q000502:physiology*; D002454:Cell Differentiation; D019070:Cell Lineage; D005260:Female; D005434:Flow Cytometry; D007381:Intermediate Filament Proteins / Q000378:metabolism; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D008648:Mesoderm / Q000378:metabolism; D051379:Mice; D008809:Mice, Inbred C3H; D032448:Myoblasts, Skeletal / Q000502:physiology*; D009203:Myocardial Infarction / Q000139:chemically induced / Q000628:therapy; D032383:Myocytes, Cardiac / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D009474:Neurons / Q000502:physiology*; D014161:Transduction, Genetic
chemical_list: D015415:Biomarkers; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin