Differentiation of hBMSC appeared to be sensitive to the exposure of very-low frequency EMF in presence of certain chemicals in the culture medium. During the period of treatment with KCl, hydrocortisone, forskolin, valporic acid and insulin the cells were exposed to EMF (50 Hz, 1 mT) for six days. RT-PCR results showed that EMF exposure significantly increased gene expression of MAP2, nestin and NF-L. Neuronal expression was also confirmed by immunocytochemistry with the neuronal marker like TUJ-1 and GFAP. Cells under the treatment with the above factors and EMF showed distinct morphology of elongated shape of neuronal cells and higher expression of neuronal marker. EMF exposure increased the intracellular Ca2+ level in the presence of the chemicals. By addition of neifedipine (L-type blocker), the intracellular calcium level was reduced in the cell exposed to EMF, which suggest that the role of L type Ca2+ channel is specifically involved in the effect mediated by EMF. The transcription factor, cAMP response element binding protein (CREB), was activated by phosphorylation within 10 min in the cells exposed by EMF without affecting activation of ERK that has been known to be a key regulator of MSC differntiation. Treatment of KN 62, a specific inhibitor for CaMKII, with the cells under EMF exposure resulted in the inhibition of the stimulatoty effect of EMF on neuronal differentiation. These results indicated that EMF exposure enhanced the level of hMSC neuronal differentiation through CaMKII signaling pathway and increasing influx of intracellular Ca2+.

Stimulation of neuronal differentiation by low frequency magnetic field on bone derived mesenchymal stem cell

248th National Meeting (2014)

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