Objective:In this study,three methods of removing microglial cells from primary astrocyte culture were compared and analyzed to explore the characteristics of different purification methods.It provides the basis for research-ers to select the appropriate primary astrocyte culture method.Methods:The cerebral cortex cells of SD neonatal rats were isolated and platted and divided into an unpurified group(control group),a traditional oscillation purification group(TO group),a cytosine β-D-arabinofuranoside(AraC)+TO group,and an AraC+L-leucine methyl ester(LME)group according to different purification methods.Cell counts were used to calculate the cell production after different purification treatments.The cell morphology of each group was observed under an optical microscope,and the proportion of glial fibrillary acidic protein(GFAP)positive cells was calculated by immunofluorescence staining.AlamarBlue HS was used to detect cell viability after passage.Results:Compared with the control group,the cell purity(P＜0.0001)and cell viability(P＜0.0001)of purified cells in the TO group were significantly increased after pas-sage,while the total number of purified cells was significantly decreased(P＜0.001).These trends were more obvious in the AraC+TO and AraC+LME group,and the cell purity(P＜0.0001 or P＜0.0001)and cell viability(P＜0.01 or P＜0.001)of purified cells in the AraC+TO and AraC+LME group after passage were significantly higher than those in the TO group.The total amount of cells obtained was significantly decreased compared with the TO group(P＜0.01 orP＜0.01).There was no significant difference between the AraC+TO and AraC+LME group.Conclusion:The culture rich in astrocytes(cell purity＞90％)was obtained by the TO method,and the cell yield was high,which could be applied to routine cell research.The highly enriched astrocyte cultures(cell purity＞99％)obtained by the AraC+TO and AraC+LME purification methods can be applied to study the precise role of astrocytes in the context of neuroinflammation.
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