(1)　The　study　of　neural　stem　cells　(NSC)　has　attracted　much　attention　in　recent　years　because　of　their　therapeutic　potential.　However,　the　problem　in　culture　and　differentiation　of　NSC　was　how　to　obtain　single　cell　suspension　that　preserves　the　function　of　NSC,　and　remove　the　debris　caused　by　mechanical　dissociation.　In　the　present　study,　we　try　to　find　a　simple　and　effective　way　to　address　the　problem,　i.e.　differential　centrifugation.　(2)　After　a　gentle　mechanical　dissociation　using　Pasteur　pipette,　the　suspension　was　first　centrifuged　at　100　g　for　5　min,　and　then　recentrifuged　at　400　g　for　6　min.　Finally,　the　two　deposits　were　resuspended　and　seeded　into　culture　flask　respectively.　The　suspension　from　the　second　deposit　was　allowed　for　further　culture　and　differentiation.　Immunofluorescence　technique　was　used　to　identify　neural　stem　cell,　neuron,　astrocyte,　and　oligodendrocyte.　(3)　After　the　second　differential　centrifugation,　single　cell　suspension　was　obtained　with　2-3　cell　clusters,　and　the　cells　not　only　grew　to　form　neurospheres,　but　also　differentiated　into　neurons,　astrocytes,　and　oligodendrocytes.　(4)　Differential　centrifugation　is　a　simple　and　effective　way　to　obtain　single　cell　suspension,　which　will　help　make　large-scale　production　of　neurodifferentiated　cells　more　effective.