小鼠脾脏来源树突状细胞的体外扩增培养(英文)

In vitro amplification and culture of mouse spleen derived dendritic cells

目的:对体外诱导小鼠脾脏单个核细胞分化成的树突状细胞(DC)的生物学表型及功能进行检测,并与骨髓来源的DC进行比较。方法:分离小鼠脾脏单个核细胞,在体外培养的条件下通过添加25 ng.ml-1的粒-单核细胞集落刺激因子和25 ng.ml-1的白细胞介素-4(IL-4)将其诱导为DC,分别从细胞形态、表型以及功能3个方面对其进行检测,并与骨髓来源的DC进行比较。结果:小鼠脾脏来源的DC经磷酸脂多糖(LPS)刺激成熟后其表面显示出明显的树枝状突起,高表达CD11c、CD86及MHC-Ⅱ类分子,并具有极强的刺激同种异基因淋巴细胞增殖的能力,其特征与骨髓来源的DC相差无几。结论:小鼠脾脏单个核细胞能够被诱导为具有正常形态、表型、功能的DC,为DC的功能研究以及进一步制备相应的肿瘤疫苗提供又一可靠来源。

Objective： To induce dendritic cells from mouse spleen mononuclear cells in vitro, and to examine the morphology, phenotype and function of the dendritic cells. Methods： Mononuclear cells isolated from mouse spleen cells were induced into dendritic cells in vitro by supplementing 25 ng·ml^-1 GM-CSF and 25 ng·ml^-1 IL-4, and then their morphology, phenotype and function were examined. Results： Mouse spleen derived DCs were induced into mature through exposure to lipopolysaccharide（LPS） and exhibited dramatic veils of cytoplasm and extensive dendrites in their surfaces with high expression of CD11 c, MHC-Ⅱ and CD86. Their capacity to stimulate primary T cell responses in mixed lymphocyte reaction （MLR） increased. The characteristics of mouse spleen derived DCs were similar to those of bone marrow DCs（BM-DCs）. Conclusion： Mouse spleen derived mononuclear cells could be induced into DCs with normal morphology, phenotype and function, which provides a new reliable source for the research of DC function and the preparation of corresponding anti-tumor vaccine.