新生SD大鼠脊髓星形胶质细胞培养

目的学习星形胶质细胞的培养方法，获得较纯的星形胶质细胞，为进一步研究打下基础。方法取出生3d内的新生SD大鼠，无菌取脊髓，剥除脑脊髓膜，剪成乳糜状后胰酶消化结合机械吹打使组织分散，筛网过滤制成单细胞悬液，差速粘附处理去除成纤维细胞得到次细胞悬液，接种用含20％FBS的高糖DMEM培养基培养，每3d换液一次。培养9～12d，待细胞汇合度达到80％时传代。取第四代细胞行GFAP免疫组化鉴定。结果成功分离培养并纯化了脊髓星形胶质细胞，经GFAP免疫组化鉴定，星型胶质细胞纯度达98．5％以上。结论细胞分离培养结合差速粘附可获得较纯的星形胶质细胞。

Culture of Astrocytes from Neonatal SD Rat Spinal Cord Tissue in Vitro

Objective To learn how to culture spinal cord astrocyte in vitro and obtain purified astrocytes to settle base for further study.Methods The spinal cord of neonatal SD rat was isolated and demeninged, mechanical methods as well as trypsin were used to make an initial cell suspension,then reduce the amount of contaminating fibroblasts by differential attachment.The secondary cell suspension was planted into 25cm^2 culture flask with HG- DMEM contain 20%FBS,the culture media was changed every 3 days,the cell passaged when it comes to appropriate 80% confluence after 9 to 12 days.The passaged cell was identified by immune stained with anti-GFAP.Result Manage to culture and purify primary spinal cord astrocyte,the cell shows satellite shape with processes,big cell body abundant cytoplasm and round or oval-shap unclear leaned to one side.Anti-GFAP immune stain identified them as target cell-astrocyte.Conclusion This method can be used to culture and purify spinal cord astrocyte.