Objective:To establish the method of culturing neural stem cells(NSCs) from rat brain. Methods:E14～16 SD embryonic rat brain and postnatal one-day(P1) rat brain were respectively chose to culture neural stem cells

and two digestive methods that simple mechanical digestion and mechanical combined with chemical digestion were used to determine the best conditions in primary culture.Immunocytochemistry and immunofluorescence were used to identify NSCs

neurons and neuroglial cells. Results:Both the primary cells from two different kinds of rat brains expressed nestin antigen and the differentiated cells expressed GFAP and MAP-2 antigen.Conclusion:The key point of primary NSCs culture was obtaining the monoplast suspension.Overdigestion led to cell injury even death. NSCs from embryonic rat brain had better proliferation ability and could form cell clones earlier and more than NSCs from newborn rat brain

but they were easier to be contaminated. Obtaining newborn rat brain and isolation NSCs from newborn rat brain were easier

but NSCs from newborn rat brain had worse proliferation ability.