人树突状细胞的体外培养和鉴定

目的:探索在体外分离培养人外周血树突状细胞的方法,同时为进一步研究树突状细胞工作奠定实验基础.方法:采用Ficoll、Precoll和Panning法分离和纯化人外周血DC,再加GM-CSF(100μg/mL)和IL-4(10μg/mL)诱生出大量树突状细胞,并进行免疫组化鉴定.结果:培养到第3天的DC扫描电镜可见DC伸出的树突状突起,呈毛刺状.培养到第7天的DC扫描电镜可见细胞胞体体积变大.这些毛刺状细胞表面有丰富的呈分叉的树突状胞质突起,某些突起形成薄片状结构,培养后的DC细胞呈S-100蛋白阳性染色.结论:人外周血树突状细胞可以在体外大量培养.

Culturing and Identifying the Human Dendritic Cells in Vitro

Objective: Explore the experiment method for cultured and identified the human dendritic cells in vitro. Simultaneously, the study would provide requirement for DC research.. Methods :Dendritic cells were isolated from human peripheral blood by Ficoll-Hypaque and Precoll, purified by the way of Panning, and cultured in the medium containing GM-CSF (granulocyte/macrophage colony- stimulating factor, 100μg/mL) and IL-4 (interleukin-4, 10μg/mL), and a large of dendritic cells were harvested. Then they were identified by immunohisto- chemical methods. Results: Dendritic cells had dendritic prominency in 3rd day after harvested viewed by scan electromicroscope. After 7th day, dendritic cells larged and had sheet-like veils or lamel-lipodia on the surface of cell. They behaved the positive staining of S-100 protein antibody. Conclusion :Dendritic cells could be isolated from human peripheral blood and cultured a large of dendritic cells in vitro.