| Budget Amount *help |
¥3,850,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥150,000)
Fiscal Year 2007: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2006: ¥3,200,000 (Direct Cost: ¥3,200,000)
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| Research Abstract |
This study was aimed to differentiate bone marrow stromal cells and neural stem cells into neurons without gene manipulation, and, moreover, to control the regeneration of neuronal circuits by electrical stimulation. We made the hypothesis that electrical stimulation, instead of gene manipulation, enable to promote the bone marrow stromal cell's differentiation into neurons. Although we tested electrical stimulating cultivation in various conditions, morphological changes into neurons could not observed. Next, electrical stimulation against the bone marrow stromal cells within culture medium containing bFGF, CNTF and Forskolin was performed. Although the expansion of neurite-like process was observed, which did not demonstrate the neuronal cell differentiation in immunohistochemistry. Electrical potential did not influence the direction of neurite expansion. Neuronal stem cells were attempted the electrical stimulation in the same way. We could not observe the neuron induced from neurosphere. Poly-L-lysin and laminin coating of the dish promote the neuronal induction from neurosphere and neurite expansion, however, which were not influenced by additional electrical stimulation. We also attempted the electrical stimulation against spinal dorsal root ganglion cells as mature neurons. Poly-L-lysin and laminin coating promoted the neurite expansion. However, velocity and direction of neurite expansion were not influenced by electrical stimulation. These results demonstrate that neurite expansion in vitro essentially needs poly-L-lysin and laminin as a substance for foothold, which suggests that enough substances for foothold connecting continuously between stem cells and native neurons are essential in the injured brain. As a subject in the future, we will investigate whether the electrical stimulation in vivo under such condition influences the velocity and direction of axonal expansion.
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