猪皮肤源树突状细胞的分离和鉴定

Isolation and Identification of Swine Skin-derived Dendritic Cells

分离出不需要体外诱导树突状细胞(DC),为研究病毒与猪DC的关系奠定基础。将刮下的猪皮肤表皮培养过夜,对所分离获得的细胞通过流式细胞术、吉姆萨染色与扫描电镜进行表型与形态鉴定。试验结果表明,通过细胞计数分离到的细胞能达到5×106个细胞,流式细胞术检测细胞CD1a/SWC3a的双阳性细胞率为82.3%;流式细胞术检测CD80/86分子,阳性细胞率为90.2%;通过吉姆萨染色和扫描电镜观察,细胞表面有大量的树突状突起,符合DC的形态特征。通过表型和形态学分析,说明已经成功分离到猪皮肤源DC,为猪病毒性疾病的免疫抑制机理研究奠定基础。

To separate dendritic cells （DC） without a requirement for in vitro propagation, this will lay a foundation for the study on the relations between the virus and swine DC. Skin layers were cut by hand with a blade and cultured overnight, and the isolated cells were acquired. The phenotype and morphology of DCs were identified by flow cytometry, Giemsa staining and scanning electron microscope. The amount of skin-derived dendritic cells by way of cell counting was 5×106. These cells exhibited 82.3% rate of positive cells of CD1a and SWC3a antigens by flow cytometry （FCM）, which was with one accord for specific molecular of DC; 90.2% rate of positive cells of CD80/86 was detected by FCM; There were more dendritic protrusions on the cells surface by way of Giemsa staining and scanning electron microscope. The skin-derived DCs of swine were isolated successfully according to the analysis of phenotype and morphology. The segregated swine skin-derived dendritic cells lay a foundation for the research on immunosuppression mechanism of viral diseases in porcine.