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目的 从人外周血中分离、培养及鉴定树突细胞 (DC)。 方法 从正常人外周血分离获得单个核细胞 ,培养 2 h后 ,取粘附细胞 ,在无血清培养基中加入不同的细胞因子 ,于培养的第 1,6 ,8天对形态、表型和功能分别进行测定 ,并在光镜、电镜下观察 DC诱导及生长情况 ,定期检测 DC的纯度与得率。 结果 体外培养 6~ 8天 ,可获得高纯度大量的 DC,较高表达 HL A- DR、CD40 、CD83和 CD86 ,能强烈激发同种异体 T淋巴细胞增殖。 结论 上述方法可以从人外周血中获得较大量的、纯度较高的 DC。
Objective To isolate, culture and identify dendritic cells (DCs) from human peripheral blood. Methods Mononuclear cells were isolated from peripheral blood of normal people. After culturing for 2 hours, adherent cells were taken and different cytokines were added in serum-free medium. The morphological, phenotypic and Function were measured, and under light microscopy, electron microscopy DC induction and growth, and regular detection of DC purity and yield. Results In vitro culture of 6 to 8 days to obtain a large number of high-purity DC, higher expression of HL-DR, CD40, CD83 and CD86, can strongly stimulate allogeneic T lymphocyte proliferation. Conclusion The above method can be obtained from human peripheral blood larger amount of high purity DC.