Abstract： Objective To study the culture and differentiation into neuron-like cells of mice amniotic fluid stem cells (mAFSCs). Methods mAFSCs separated from the amniocyte were cultured and sub-cultured. The proliferation and growth characteristics were observed in primary and passage cultures. The cell-surface antigens and cell cycle of mAFSCs were analyzed with flowcytometry. After mAFSCs were induced to differentiate into neuron-like cells, neuron-β-tubulin was detected by immunofluorescence. Results The primitive stem cells from mouse amniotic fluid could reach above 80% in 7 to 10 days, and then the number of the cells was 3.5×105-4.0×105. After more than 3 passages, the appearance of mAFSCs was spindle-shaped, and the alignment of the cells showed bars. The mAFSCs had a good viability and proliferation in vitro. After cultured up to 90 days, the cell appearance changed and capability of proliferating decreased in mAFSCs. The cell exterior antigen was analyzed by flow cytometry,and Sac-1 ,CD29 were positive, CD11b, CD34, CD45 were negative. The mAFSCs were differentiated into neuron-like cells after induction by neuron inducing medium for 3 days, and some of them expressed the specific antigen neuron-β-tubulin. Conclusion The cell viability and proliferation of mAFSCs are good, and they also have the ability to directional differentiation, which may provide a new stem cell for cell therapy in mouse research.