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《Neural Injury and Functional Reconstruction》 2014-06
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Primary Culture and Purification of Microglia from Rat Spinal Cord

CHEN Xue;ZHANG Ting;LI Cai-hong;WANG Wei;LUO Xiang;YU Zhi-yuan;Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology;  
Objective: To establish a simple and highly efficient primary culture method for microglia from rat spinal cord. Methods: Mixed glial cells were obtained from the spinal cord of newborn SD rats. On day 3, the culture medium was changed until the cultured microgila became fused on day 10. The microglia were purified using shocking methods(37 ℃, 180 rpm, 1~2 h) with different adhesion time(20~40 min). The isolated and purified microglia were identified with Iba1 and CD11b after 24 h. Results: The rat spinal cord microglia were successfully isolated and purified. The cultured microglia presented round or fusiform, highly refractive morphology. Iba1 and CD11b fluorescence tests showed that the purity of microglia was more than 95%. Conclusion: Using nutrition deprivation, Shock and different adhesion time methods, a primary spinal cord microglia culture method with high yield and purity was established, providing a useful tool for studying rat spinal cord microglia in vitro.
【Fund】: 国家自然科学基金(No.81000521 81030021)
【CateGory Index】: R741.02
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