Objective To establish a method for the in-vitro culture of astrocytes from cerebral cortex of neonatal rats.Methods Astrocytes were isolated from cerebral cortex of the neonatal SD rats aged 24 hours,and then were cultured in the high-glucose DMEM medium with 10% fetal bovine serum added.The morphological features of astrocytes were examined under inverted microscope.The growth of the third generation of astrocytes was observed with counting plate method and methyl thiazolyl tetrazolium (MTT) assay.Results The astrocytes from cerebral cortex of neonatal rats grew well,the astrocytes showed typical morphological features,and no other kinds of cells was found.Conclusion A simple and practical in-vitro culture method for rats astrocytes is established.