Effect of brainderived neurotrophic factor in neural differentiation of mouse embryonic stem cells and neural precursor cells
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abstract
Embryonic stem (ES) cells, which have pluripotency in differentiation, have been considered as an excellent source for cell replacement therapy of damaged tissues. Although neurotrophic factors induce neural differentiation in neural precursor cells, their actions in ES cells are not clear. This study was designated to examine the effect of brain-derived neurotrophic factor (BDNF) on neural differentiation in mouse ES cells. Undifferentiated mouse ES cells were maintained at suspension culture in the presence of leukemia inhibitory factor (LIF), and neural precursor cells were isolated from the subventricular zone (SVZ) of the 2-3 day old mice brain. Oct-4 was expressed in undifferentiated ES cells and neural precursor cells, of which expression was markedly decreased by induction of differentiation. RT-PCR analysis showed that neural differentiation of ES cells induced expression of various neural markers, and that BDNF increased their expression. Western blot analysis demonstrated that BDNF induced expression of synaptophysin, a neuronal marker, and inhibited expression of glial fibrillary acidic protein, a glial marker. Undifferentiated ES cells expressed TrkA, TrkB and TrkC, of which expressions were increased during differentiation. Neural differentiation increased their expression, and BDNF further increased them. The treatment of BDNF during differentiation decreased expression of Notch1. These results indicated that BDNF induces neural differentiation and inhibits glial differentiation in ES cells as well as neuronal precursor cells.
