大鼠脊髓星形胶质细胞的体外培养与鉴定

目的:探讨大鼠脊髓组织星形胶质细胞体外培养、纯化和鉴定的方法。方法:于无菌条件下取新生1~2 d的SD大鼠脊髓组织,采用机械吹打及胰酶消化法制成细胞悬液,通过差速贴壁和恒温摇床震荡去除杂细胞,进行培养。用胶质纤维酸性蛋白(GFAP)免疫细胞化学染色对所培养的细胞进行鉴定。结果:分离培养的细胞具有典型的星形胶质细胞的形态,传至第3代细胞纯度达95%以上。结论:成功建立了大鼠脊髓星形胶质细胞体外培养方法,为脊髓星形胶质细胞的实验研究奠定基础。

Culture and Identification of Rat Spinal Cord Astrocyte in Vitro

Objective：To explore the methods of purification and identification of astrocytes in vitro rat spinal cord tissue culture.Method：Rats aged 1 to 2 day were used in the experiment.Spinal cord was dissected under sterile operation and cells suspension through the mechanical method and enzymatic digestion.Differential attachment and orbital shaking bed were used to remove contaminating cells.Finally the passaged cell was identified with immunofluorescence staining against glial fibrillary acidic protein（GFAP）.Result：The rat spinal cord astrocytes were successfully isolated and the proportion of astrocyte was beyond 95%in the third generation cells.Conclusion：The cultivation method of astrocyte from rat spinal cord is established with high purity to provide basal experimental models for spinal cord astrocyte research.