An improved culture method of astrocytes in cerebral cortical of rats
Pages: 349-353
Year: 2017 Issue: 3
Journal: Chinese Journal of Neuroanatomy
Abstract: Objective:To improve the culture method of astrocytes in the cerebral cortex of rats,and obtain high purity astrocytes and used in neuroscience research.Methods:Newborn rats (2 ～3 days age) were cut off head after 75％ alcohol disinfection,removed the cerebral cortex on the ice,then put into the cold D-Hanks solution.The meninges and blood vessels were removed,then cut up and digested by 0.25％ pancreatic enzyme to separate cells,paving 0.015％ Poly-D-Lysine solution in the bottle before cells vaccination by the density of 1 × 105 cells/m1.In order to get high purity astrocytes,cell purification used the method of "differential adhension","gradient of serum" and "cross in hand".Cells purity was identified by immunofluorescence cell staining of glial fibrillar acidic protein (GFAP),which is the specific protein expressed in astrocytes.Results:Astrocytes purity is more than 98％.Cells can be multiple batches,cell proliferation was fast and grow well.Conclusion:This improved method of the primary culture of astrocyte cells is simple operation,high purity,can satisfy the experimental requirements of high purity astrocytes culture for various kinds of neuroscience research,suitable for promotion.
Year: 2017 Issue: 3
Journal: Chinese Journal of Neuroanatomy
Abstract: Objective:To improve the culture method of astrocytes in the cerebral cortex of rats,and obtain high purity astrocytes and used in neuroscience research.Methods:Newborn rats (2 ～3 days age) were cut off head after 75％ alcohol disinfection,removed the cerebral cortex on the ice,then put into the cold D-Hanks solution.The meninges and blood vessels were removed,then cut up and digested by 0.25％ pancreatic enzyme to separate cells,paving 0.015％ Poly-D-Lysine solution in the bottle before cells vaccination by the density of 1 × 105 cells/m1.In order to get high purity astrocytes,cell purification used the method of "differential adhension","gradient of serum" and "cross in hand".Cells purity was identified by immunofluorescence cell staining of glial fibrillar acidic protein (GFAP),which is the specific protein expressed in astrocytes.Results:Astrocytes purity is more than 98％.Cells can be multiple batches,cell proliferation was fast and grow well.Conclusion:This improved method of the primary culture of astrocyte cells is simple operation,high purity,can satisfy the experimental requirements of high purity astrocytes culture for various kinds of neuroscience research,suitable for promotion.
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