Objective: To isolate neural stem cells from the hippocampus of neonatal rats and to induce them to differentiate into cholinergic neurons.Methods: A multipotent cell line derived from the hippocampus of neonatal rats which had the ability to form clones was incubated in serum-free DMEM/F12 media added with 20ng/ml b-FGF, B27. Immunocytochemistry was used to detect Nestin antigen of the cell clone and specific antigen of neuron(Tujl), astrocyte(GFAP), and oligodendrocyte(Galc) after differentiation of the neural stem cells. Embryonic chick skeletal muscle extract was used to induce the differentiation of the neural stem cells into cholinergic neurons.Result: The cell line isolated from the hippocampus of neonatal rats expressed Nestin antigen and had the potential to form clones and differentiate into neurons, astrocytes and oligodendrocytes. Embryonic chick skeletal muscle extract could induce 9.6% of the the isolated cell line to differentiate into cholinergic neurons compared with 3.9% in controls.Conclusion: The cell line, which expressed Nestin antigen was a multipotent cell line and had the ability to self-renew. It is believed that it contained stem cells of the CNS. These neural stem cells could be induced to differentiate into cholinergic neurons by using embryonic chick skeletal muscle extract.
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