Isolation, Cultivation And Identification Of Neural Stem Cells From Human Embryonic Cerebral Tissue

Objective: To study the growth characterization differentiation and the cultivation condition of neural stem cells from human embryonic cerebral tissue in vitro for further related reseach.Method: To isolate forebrain and hindbrain of human fetuses about 9 weeks and 12 weeks from spontaneous abortion. Neural stem cells were cultured in special culture medium and were compared with growth characterization.The differentiation from these neural stem cells was investigated in both a fetal bovine serum-containing and a non-supplemented,serum-free basal medium.Immunofluorescence cyto -chemical method was used to detect Nestin antigen of neural stem cells and specific antigen of mature neural cells after the neural stem cells differiented.Results: The neural stem cells with positive expression of Nestin were isolated from forebrain and hindbrain of human fetuses about 9 weeks and 12 weeks. The cells differentiated from these neural stem cells expressed specific antigen of neuron ,astrocytes and oligodendrocytes inboth a fetal bovine serum-containing and a non-supplemented,serum-free basal medium. The ratio of cell clone formation from human embryonic cerebral tissue about 9 weeks is higher than that of 12 weeks in primary culture. Under the existence of special medium,the differentiation of neural stem cells from hindbrain is easier than that from forebrain .Conclusion: The neural stem cells can be isolated from human embryonic cerebral tissue . The ratio of cell clone formation from smaller gestational age is higher than that from elder gestational age .The differentiation of neural stem cells from hindbrain is easier than that from forebrain.