Objective    To establish experimental cell models of myogenic cells

adipocytes

endothelial cells and neurocytes differentiated from murine embryonic mesenchymal stem cell line C3H/10T1/2.      Methods     Myogenic and adipogenic differentiation of C3H/10T1/2 cells were induced by 5-azacytidine

angiogenic differentiation by vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF)

and neurogenic differentiation by bFGF

β-mercaptoethanol (β-ME) and DMSO. The differentiated cells were identified by observing cell morphology

Oil Red O staining and immunohistochemical staining to detect endothelial cell surface marker CD31 and neuron-specific enolase (NSE).     Results    C3H/10T1/2 cells were morphologically elongated at the 10th day

and myotube-like cells appeared at the 17th day after 5-azacytidine induction. Meanwhile

adipose globelets were observed in a small proportion of cells at the 17th day

and Oil Red O staining confirmed adipose accumulation in the cytoplasm at the 25th day after 5-azacytidine induction. Cobble stone-like structure was observed since 5th day after inducement of VEGF and bFGF

and at the 8th day the induced cells exhibited typical CD31 positive staining. After treated with bFGF

β-ME in combination with DMSO

cytoplasm retracted towards the nucleus and the cell processes prolonged at the 3th day. Immunohistochemical staining showed that differentiated cells expressed NSE at the 15th day.      Conclusion    C3H/10T1/2 cells have the potential of multilineage differentiation and can be used as an alternative source of mesenchymal stem cells. Growth factor-induced endothelial differentiation of C3H/10T1/2 cells was established.