Optimization and methods of culture in vitro of astrocytes from cerebral cortical mice
Objective To explore and optimize the in vitro primary culture method of astrocytes in neonatal mouse cerebral cortex, which provides a better solution for the in vitro culture of astrocytes.Methods In order to opti-mize the in vitro culture method of mouse cerebral cortex astrocytes, 3-day-old C57BL/6J mouse cerebral cortex tis-sues were taken, meninges and blood vessels were removed, digested by pancreatic enzymes and centrifuged, and high-glucose dulbecco's modified eagle medium (DMEM) was added to form cell suspension, which was purified by differential adhesion method, cross hand method and constant temperature shaking method.The cells were inoc-ulated in poly-D-lysine-coated culture bottles with different culture densities, and the purity of astrocytes was deter-mined by morphological observation and immunofluorescence staining.Results The cells were inoculated at a den-sity of 5 × 106 cells per bottle with good effect and high activity.The purity of astrocytes reached 99％ by using high sugar DMEM medium combined with differential adhesion method, cross hand method and constant temperature shaking method.Conclusion The primary culture method of astrocytes in mouse cerebral cortex is successfully es-tablished and optimized.
astrocytecerebral cortexprimary culturecell purificationglial fibrillary acidic protein
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