Neural Stem Cells was Induced to Differentiate into Cholinergic Neurons in Vitro

The cholinergic-inducing effect of BMP4 on isolated and cultivated rat’s cerebral neural stem cells(NSCs) was examined.NSCs which were isolated from two month’s old rat’s brain region like hippocampus and striatum were cultivated in a medium containing EGF and bFGF, and were identified with morphological character by microscope and nestin immunocytochemistry test.After 24 hours, half NSCs were cultivated with a BMP4-added medium as a experimental group instead of the primary medium,while the an other half NSCs being cultivated with the primary medium as a control group.After 8 days the expression of choline acetyltransferase(ChAT) of the cultivated cells was observated by indirect immunofluorescence test. Results showed that more positive cells were found in the experimental group, and the fluorescence intensity were stronger; while less positive cells were found in the control group, and the fluorescence intensity was weaker.The differentiational efficiency of the NSCs was examined by FITC-labelled Flow Cytometry.The results showed that about 16% cells of the experimental group appeared ChAT-positive, while that of control group only 7%.So BMP4 may have the function of inducing NSCs to differentiate into neurons with cholinergic characteristic.