人外周血树突状细胞的体外培养扩增

Proliferation of human dendritic cells from monocyte in vitro

目的： 在体外从人外周血中培养扩增树突状细胞。方法：从正常人外周血分离PBMC，经两次孵育粘附，弃去悬浮细胞后加细胞因子（GM-CSF，IL-4）和培养液进行培养，并对培养过程进行观察鉴定。结果：培养1周后即可培养出典型的树突状细胞，经FITC-CD1a单抗标记后用流式细胞仪检测DC的CD1a表达率为 65%，并能刺激同种淋巴细胞增殖反应。结论：利用GM-CSF和 IL-4可以从人外周血中扩增出大量DC，可为后续研究做准备。

Objective: To proliferate dendritic cells from human peripheral blood. Methods: PBMC isolated from healthy human peripheral blood were incubated for 3 hours twice at 37℃ in humidified 5%CO2. After 3 hours, nonadherent cells were gently removed, then the adherent cells were cultured at 37℃ in humidified 5%CO2 in air,in medium with GM-CSF and IL-4 for more than 7 days. Finally, DCs were collected and identified in morphology，function and specific symbol of cell surface. Results: After 7 days, typical dendritic cells appeared and gradually became larger and more typical.By using flow cytometric cell surface marker analysis ,the level of expressing CD1a was 65%. DCs could also induce proliferation of allogeneic lymphocytes. Conclusion: A large amount of DCs could be induced from peripheral blood monocyte by adding GM-CSF and IL-4.