Abstract: ［Abstract］〓Objective〖HTSS〗〓To establish a simple method for isolation and culture of mouse neural stem cells(NSCs) from ICR mice in vitro.
〖HTH〗〖WTHZ〗Methods〖HTSS〗〓The brain tissues of ICR mice were isolated and cultured in serumfree medium. The morphology of cultured cells was observed with an inverted microscope. The specific markers, nestin, glial fibrillary acidic protein(GFAP) and neuronal specific enolase(NSE) were detected by immunocytochemical staining to identify cells. The growth curve of the cells was plotted by CCK8 assay.
〖HTH〗〖WTHZ〗Results〖HTSS〗〓The cells isolated from the embryonic brain tissue of ICR mice were amplified in the form of neurospheres in serumfree medium, and could be passaged continuously after digestion. The growth curve showed that the cell doubling time was 2 d. The percentage of nestin positive cells was(72.0±5.5)% in neurospheres. After induced differentiation, two types of cells were observed: one has long and thick projection interlaced into a net, and is GFAP positive; the other is polygonal, with a number of slender projections, and is NSE positive.
〖HTH〗〖WTHZ〗Conclusion〖HTSS〗〓The NSCs cultured by this method has high purity, proliferate rapidly, and have the potential of multidifferentiation. Our method is worth popularizing and applying.

Key words: neural stem cells, cell culture techniques, mice