Isolation and characterization of bone marrow-derived mesenchymal progenitor cells with myogenic and neuronal properties.
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.
Shiota
Mitsutaka
M
Department of Pediatrics, Graduate School of Medicine, Kyoto University, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Heike
Toshio
T
Haruyama
Munetada
M
Baba
Shiro
S
Tsuchiya
Atsunori
A
Fujino
Hisanori
H
Kobayashi
Hirohiko
H
Kato
Takeo
T
Umeda
Katsutsugu
K
Yoshimoto
Momoko
M
Nakahata
Tatsutoshi
T
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
D015415:Biomarkers; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin