小鼠骨髓源树突状细胞的扩增及鉴定

Generation and identification of dendritic cell from murine bone marrow

目的建立小鼠骨髓源树突状细胞的体外扩增方法并研究树突状细胞的相关特征。方法根据不同时期树突状细胞的粘附性质不同,设计简便的树突状细胞的体外扩增和纯化方法,并利用瑞氏染色和流式细胞仪(FACS)鉴定其生物学特征。结果体外培养24 h后,可见增殖性细胞集落,3 d后集落增多明显,体外培养7 d,收集悬浮细胞即为DC。光镜显示细胞表面不规则,呈树突状突起,瑞氏染色嗜碱性。流式细胞鉴定为髓系DC,高表达MHCⅡ类分子及共刺激分子(CD40、CD80、CD83、CD86)。结论此简便方法制备的树突状细胞具有髓系树突状细胞的生物学特性,及较高的纯度,可广泛的应用于临床及实验研究。

Objective To create a method of generate dendritic cell （DC） from murine bone marrow, and explore the correlative character of DC. Methods According to different adhesiveness of DC in vary period, a simple method was used to generate and purificate DC, and their biology character were identified by Wright staining and FACS. Results The clusters of cell were saw after 24 hours cell cultured, and increased significantly after 3 days. After 7 days, the suspension cells were collected, which were DCs. The branching structures on the Dendritic Cell were visualized under the inverted light microscope. The bias nucleus of DC was basophilic after Wright staining. These DCs belonged to myeloid lineage DC with high expression of MHC Ⅱ and co-stimulatory molecules （ CD40, CD83, CD80, CD86）. Conclusion The cultured murine bone marrow DC have the same characteristics as myeloid DC and are widely used for clinical study and Experiment.