The diferentiation of neuronal cells from Mouse embryonic stem cells
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Date
2014
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Abstract
With new technologies emerging today, the importance of stem cells in the cell therapy of nervous system diseases is supportedby recent studies. Therefore, the development of neuronal cell diferentiation protocols from stem cells is of great importance. Inour study, the diferentiation of neuronal and neuroglial cells from mouse embryonic stem (ES) cell line and their analysis withneuronal cell markers are aimed. Mouse ES cells were diferentiated to neurogenic series cells by adding N2 and bFGF to the culturemedium on coated Fibronectin dishes. For the identification of diferentiated cells, they were evaluated by light microscopy usingimmunhistochemistry techniques and by electron microscopy. Indirect immunohistochemical staining method was performedwith SSEA-1 (mouse embriyonic stem cells marker), Nestin (neural precursor cells marker), βIII-Tubulin (neuronal cells marker),MAP-2 (neuronal cells marker), GFAP (astrocyte marker), and O4 (oligodendrocyte marker). After 1 week of diferentiation of cells,immunoreactivities of SSEA-1 and Nestin were detected to be negative and moderate, respectively. After 2 weeks culture time, thediferentiation was still continuing and especially positive immunoreactivities of β-III Tubulin and MAP-2 and weak immunoreactivitiesof O4 and GFAP were supported neuronal diferentiation. In conclusion, our results suggest that neuronal cell derived from mouse EScells were diferentiated particularly to neuron using N2+bFGF+fibronectin culture condition. Therefore, these diferentiated cells maybe used as a treatment method in degenerative diseases of the nervous system.