Abstract:

Objective： To observe the morphology and growth pattern of the neural stem cells neural stem cells（NSCs） in hippocampus tissue， and to provide the basis for the extraction and culture of the NSCs. Methods The NSCs were isolated from hippocampus tissue of the newborn SD rats.The morphology of the NSCs in hippocampus tissue was observed； flow cytometry was used to detect the purities of the NSCs in hippocampus tissue； immunofluorescence method was used to detect the expressions of Nestin and EdU proteins in the NSCs； the proliferative activities of the NSCs were analyzed by cell counting and CCK-8 methods. Results On the 0 day of culture，the NSCs in hippocampus tissue were suspended in the culture medium with the form of single cells；on the 2nd day of culture，the NSCs in hippocampus tissue suspended in the culture medium and began to gather into the cell clumps with different sizes and irregular shapes；On the 8th day of culture，the neurospheres showed round or oval neurosphere with different sizes， and showed clear boundary without synapse.The purity of the NSCs in hippocampus tissue was 76.50%－85.40% On the 8th day of culture，the positive expression of Nestin in the cytoplasm of the NSCs was green，and the positive expression of EdU in the nucleus was red.The neurospheres were composed of the Nestin and EdU co-staining cells. the NSCs were at logarithmic growth period，and the proliferation abilities were high when cultured for 5－11 d. Conclusion The proliferation ability of the NSCs is high；the purity of the NSCs is high on the 8th day of culture； the proliferation ability of the NSCs is high when cultured for 5－11 d.

Key words: Neural stem cell, Hippocampus tissue, Proliferative activity, Growth curve, Immunofluorescence