关键词: 细胞, 培养的, 诱导, 共刺激分子

Abstract: Objective To induce and culture purity dendritic cells (DCs) of different maturation phase from human peripheral blood monocytes in vitro. Methods The monocytes were acquired from human peripheral blood mononuclear cells（PBMC）by adhesion method.The first step was to work on a 7-day culture of monocytes in medium supplement with GM-CSF and IL-4, and the cells were actually immature. In the second step,the cells were cultured in the medium supplement with GM-CSF and TNF-α,and cultured until the 14th day. Mature DCs were obtained. Cells harvested were identified for their morphology by microscope, properties of DCs detected by FCM, function with MTT method. Results Immature dendritic cells expressed CD1a molecules and costimulating molecules in middle level, HLA-DR in high level. In maturation phase DCs expressed costimulating molecules, HLA-DR molecules, CD83, and CD25 highly. These DCs could stimulate proliferation of allogenic T lymphocytes and increase killing ability of CTL activated by DCs. Conclusion The method to obtain DCs of different maturation phases from human peripheral blood monocytes is successfully set up. A number of DCs with high purity are obtained.

Key words: cells, cultured, induction, costimulating molecules