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Reevaluation of in vitro differentiation protocols for bone marrow stromal cells: disruption of actin cytoskeleton induces rapid morphological changes and mimics neuronal phenotype
Journal article   Peer reviewed

Reevaluation of in vitro differentiation protocols for bone marrow stromal cells: disruption of actin cytoskeleton induces rapid morphological changes and mimics neuronal phenotype

Birgit Neuhuber, Gianluca Gallo, Linda Howard, Lisa Kostura, Alastair Mackay and Itzhak Fischer
Journal of neuroscience research, v 77(2)
15 Jul 2004
PMID: 15211586

Abstract

Chondrogenesis - drug effects Osteogenesis - physiology Actins - metabolism Gene Expression Regulation, Developmental - genetics Adipocytes - drug effects Cell Lineage - drug effects Fibroblasts - ultrastructure Neurons - ultrastructure Actin Cytoskeleton - drug effects Neurites - ultrastructure Nerve Tissue Proteins - biosynthesis Stromal Cells - drug effects Adipocytes - ultrastructure Bone Marrow Cells - drug effects Neurons - metabolism Neurons - drug effects Cell Differentiation - physiology Neurites - drug effects Fibroblasts - metabolism Embryonic Induction - genetics Actin Cytoskeleton - metabolism Osteogenesis - drug effects Stromal Cells - metabolism Cells, Cultured Gene Expression Regulation, Developmental - drug effects Rats Neurites - metabolism Nerve Tissue Proteins - drug effects Bone Marrow Cells - ultrastructure Cell Lineage - physiology Chondrogenesis - physiology Phenotype Stromal Cells - ultrastructure Animals Cell Differentiation - drug effects Adipocytes - metabolism Fibroblasts - drug effects Culture Media - pharmacology Biomarkers Growth Substances - pharmacology Actin Cytoskeleton - ultrastructure Bone Marrow Cells - metabolism Embryonic Induction - drug effects
Bone marrow stromal cells (MSC), which represent a population of multipotential mesenchymal stem cells, have been reported to undergo rapid and robust transformation into neuron-like phenotypes in vitro following treatment with chemical induction medium including dimethyl sulfoxide (DMSO; Woodbury et al. [2002] J. Neurosci. Res. 96:908). In this study, we confirmed the ability of cultured rat MSC to undergo in vitro osteogenesis, chondrogenesis, and adipogenesis, demonstrating differentiation of these cells to three mesenchymal cell fates. We then evaluated the potential for in vitro neuronal differentiation of these MSC, finding that changes in morphology upon addition of the chemical induction medium were caused by rapid disruption of the actin cytoskeleton. Retraction of the cytoplasm left behind long processes, which, although strikingly resembling neurites, showed essentially no motility and no further elaboration during time-lapse studies. Similar neurite-like processes were induced by treating MSC with DMSO only or with actin filament-depolymerizing agents. Although process formation was accompanied by rapid expression of some neuronal and glial markers, the absence of other essential neuronal proteins pointed toward aberrantly induced gene expression rather than toward a sequence of gene expression as is required for neurogenesis. Moreover, rat dermal fibroblasts responded to neuronal induction by forming similar processes and expressing similar markers. These studies do not rule out the possibility that MSC can differentiate into neurons; however, we do want to caution that in vitro differentiation protocols may have unexpected, misleading effects. A dissection of molecular signaling and commitment events may be necessary to verify the ability of MSC transdifferentiation to neuronal lineages.

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Neurosciences
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