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小鼠脾脏来源树突状细胞的体外扩增培养
作者:黎辉 曹宇皎 黄军华  
单位:河北任丘市华北石油总医院
关键词:脾脏 骨髓 树突状细胞 诱导 
分类号:
出版年·卷·期(页码):2010·29·第五期(481-484)
摘要:

背景:不同来源的树突状细胞具有不同的生物学特点。由于体外诱导脾脏来源树突状细胞的产量相对较低,因此相关研究也比较少。目的:体外诱导小鼠脾脏单个核细胞分化为树突状细胞,对其生物学表型以及功能进行检测,并与骨髓来源的树突状细胞进行比较。设计、时间和的点:基础实验,于2005-9/2006-5在贵阳医学院干细胞研究中心完成。材料:BALB/c及129小鼠购自贵阳医学院动物实验中心,提取脾脏和骨髓单个核细胞。实验过程中对动物处置符合动物论理学标准。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25ng/ml的粒-单核细胞集落刺激因子和25ng/ml的白细胞介素-4将其诱导为树突状细胞,分别从细胞形态、表型以及功能三个方面对其进行检测,并与骨髓来源的树突状细胞进行比较。主要观察指标:树突状细胞的形态学特征、表型以及刺激同种异基因淋巴细胞增殖的能力。结果:小鼠脾脏来源的树突状细胞经磷酸脂多糖(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树突状细胞相差无几。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、功能的树突状细胞,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。

BACKGROUND: The dendritic cell derived from different source has different biologic characteristic. As the yield of DC induced from spleen is lower, so the relative research is few. OBJECTIVE: This research was to induce dendritic cells from mice spleen mononuclear cells in vitro, then examined from their morphology, phenotype and function, and compared it with bone marrow derived DC. DESIGN, TIME AND SETTING: A fundamental study. The research was performed at The center of stem cell, Guiyang Medical College from September 2005 to June 2006. METERIALS:BALB/c and 129 mice were purchased from Animal center of Guiyang Medical College. Distill spleen and bone marrow mononuclear cells. Animal intervention met animal ethical standard. METHOD: mononuclear cells isolated from mice spleen cells were induced into dendritic cells in vitro by cultured in 25ng/ml GM-CSF and 25ng/ml IL-4,and then examined from their morphology, phenotype and function. MAIN OUTCOME MEASURES: The morphologic characteristic, surface marker of DC, and the capacity to stimulate primary T cell response. RESULTS: Mice spleen derived DC were induced into mature through exposure to lipopolysaccharide (LPS) and exhibit dramatic veils of cytoplasm and extensive dendrites in their surfaces, high express CD11c, MHC-Ⅱ, CD86, their capacity to stimulate primary T cell responses in mixed leukocyte reaction (MLR) would be shown strongly. The characteristics of Mice spleen derived DC were similar with BM-DC. CONCLUSION: Mice spleen derived mononuclear cells could be induced into DC with normal morphology, phenotype and function, provide a new reliable source for the functional research of DC and next produce corresponding anti-tumor vaccine

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