Objective: To isolate, cultivate and identify the neural stem cells from the cortex of embryo mouse in vitro for the further related research.
Methods: Cerebral hemisphere of mouse fetuses were taken out. Suspension of cerebral cells was made. Neural stem cells were identified by immunofluorescent cytochemistry to the primary culture. Neural stem cells were isolated and cultured in special culture medium using single-cell clone culture technique. After the stem cells were stably cultured successively for 20 generations, nestin, glial fibrillary acidic protein, beta-tublin, and galactosidase (Galc) were detected by indirect immunofluorescence cytochemistry.
Results: The stem cells isolated from the cerebral hemisphere of mouse embryo with positive expression of nestin could be passaged in vitro for at least 20 generations. Inoculated on the coverglass with poly-1-ornithine and cultured in medium containing serum, the stem cells were induced to differentiate. The cells differentiated from these stem cells showed green fluorescence by GFAP fluorescent staining, red fluorescence by beta-tublin fluorescent staining, or green fluorescence by Galc fluorescent staining.
Conclusion: The stem cells from cortex of mouse embryo can be successfully isolated and cultured in vitro. The stem cells can be passaged in series and possessed the potential of multi-directional differentiation and express the specific antigens of neuron, astrocytes, and oligodendrocytes. Thus they can be used for further experimental study.