Gene Profiles in the Early Stage of Neuronal Differentiation of Mouse Bone Marrow Stromal Cells Induced by Basic Fibroblast Growth Factor

Figure 3

Neuronal differentiation induced by bFGF. (a) Morphological observation of BMSCs subjected to 10 ng/ml bFGF for 3 and 6 days. Typical signs of a developing neuron, such as cell body shrinkage, axonal elongation, and neurite outgrowth, were observed. The distinctive observation is heightened at day 6 postadministration. (b) RT-PCR representation of marked increase in genes regulating neuronal development and maturation as evidently shown by increase of ChAT, GAP43, NSE, and Tau. (c) Western blotting results supported the same observation of increased neuronal gene expression at protein levels which strengthened the deduction of a neuronal development and eventual maturation. (d) The electrophysiological properties of these differentiated cells were examined by patch clamping method at day 6 post-bFGF administration. (e) The expressions of SERT, MAP2, and β-tubulin were determined by qRT-PCR at day 6 post-bFGF administration. Data of qRT-PCR indicate the of three experiments. (f) Distribution of neuronal differentiation markers. Immunofluorescence staining using specific antibodies against GAP43, NSE, Tau, MAP2, and β3-tubulin was performed in BMSCs with bFGF treatment for 3 days. NC: negative control. Images are representatives of at least three experiments.

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