ABSTRACT
Objective To isolate and identify the cells from the cerebral cortex,cerebellar cortex,hippocampus,sub-ventricle region,brain stem and the spinal cord of neonatal rats and observe their growth morphology in vitro.Methods The cells were incubated from different regions of the CNS of neonatal rats by using DMEM/F12 media added with 10% bovine serum,and their growth morphology was observed by using inverted phase contrast microscope;then in the 10th day after incubation cells were fixed and immunocytochemical method was used to detect specific NSE antigen of the neurons and specific GFAP antigen of the astrocytes.Results Both neurons and astrocytes were studied in each region and they bloomed in the 10th day after incubation.Neurons had big triangular or ellipsoidal cell bodies surrounded with a halo and had robust nervous process which interlaced each other around cell bodies.The astrocytes had an ellipsoidal nucleus located at one side of the cell body and they had abundant processes branching profusely.Conclusion A method of culturing cells from different regions of the CNS of neonatal rats was described.A comparative morphology study of their growth was made and neurons and astrocytes of all the regions studied were identified.
ABSTRACT
The present study aims to isolate neural stem cells from neonatal rat hippocampus and induce them to differentiate into cholinergic neurons. A multipotent cell line derived from the hippocampi of neonatal rats which had the ability to form clones was incubated in serum-free DMEM/F12 medium added with 20ng/ml basic fibroblast growth factor (bFGF) and B27. After differentiation of the neural stem cells, immunocytochemistry was used to detect nestin, the antigen of the cell clone, and β-tubulin (Tuj 1 ), glial fibrillary acidic protein (GFAP) and galactocerebroside (Galc), the markers specific for neurons, astrocytes and oligodendrocytes, respectively. Embryonic chick skeletal muscle extract was used to induce the differentiation of the neural stem cells into cholinergic neurons. The results showed that the cell line isolated from the hippocampi of neonatal rats expressed nestin and had the potential to form clones and differentiate into neurons, astrocytes and oligodendrocytes. Embryonic chick skeletal muscle extract can induce 9.6% of the isolated cell line to differentiate into cholinergic neurons compared with 3.9% in controls. These findings suggested that the cell line, which expressed nestin antigen, was a multipotent cell line capable of self-renewing, and was believed to contain stem cells of the CNS. These neural stem cells can be induced to differentiate into cholinergic neurons by using embryonic chick skeletal muscle extract.