摘要
目的:证实体外诱导分化的人树突状细胞(DC)表面高效表达树突状细胞特异性细胞间黏附分子-3结合非整合素因子(DC-SIGN),为进一步研究其在丙型肝炎病毒传播中的作用奠定基础。方法:F icoll密度梯度离心法分离外周血单个核细胞(PBMC),黏附2 h后接种于完全培养液中,并加入rhGM-CSF、rhIL-4刺激分化,共培养7天,光镜及扫描电镜观察细胞形态,以DC表面特异性标志DC-SIGN的特异性抗体进行荧光染色,检测DC-SIGN的表达。结果:经诱导产生的细胞具有典型的DC形态特征,高表达DC特异性标志物DC-SIGN。结论:GM-CSF与IL-4联合刺激PBMC可分化为DC,且高表达DC-SIGN,为后续研究DC-SIGN奠定了基础。
Objective : To determine the expression of DC-SIGN on generated dendritic cells from human peripheral blood. Methods: Peripheral blood monocytes (PBMC) were isolated from blood of healthy individuals by ficoll sedimentation according to standard procedures. Monocytes were purified from PBMC by a 2 h adherence step at 37℃ in complete medium. Nonadherent cells were washed off with PBS and the remaining adherent cells immediately subjected to the DC differentiation protocol. Monocytes resuspended at complete medium containing GM-CSF and IL-4. Cells were cultured for 7 days, with cytokine addition every second day, to obtain a population of immature DC. Characteristics of the cultured cells were observed by LM and SEM, and the expression of DC-SIGN was detected by immunofluorescence (IF). Results: After 7 days'culture, typical dendritic cells appeared and the level of DC-SIGNg expression was high. Conclusion : DC with high expression of DC-SIGN could be induced from peripheral blood monocyte in complete medium containing GM-CSF and IL-4.
出处
《医学研究生学报》
CAS
2005年第12期1066-1068,i0018,共4页
Journal of Medical Postgraduates
基金
国家自然科学基金资助项目(批准号:20170822)