人外周血树突细胞的分离与提纯

Isolation and Purification of Dendritic Cells from Human Peripheral Blood

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\ Proliferation and isolation of dendritic cells from human peripheral blood.\ Methods\ Monocytes isolated from human peripheral blood, culture for 2 hours.\ The adherent cells were cultured with different cytokines respectively in serum\|free culture.\ The cells were examined by phenotype/FACS for 1,6 and 8 days respectively,and the cell growth was inspected under a light microscope and electronmicroscope at regular interval and determined function assay.\ Results\ During 6 days to 8 days, a large number of DC with high purity were generated.\ DC expressed HLA\|DR,CD\-\{40\},CD\-\{86\} and CD\-\{83\} costimulating moleculars highly on cell surface and could stimulate proliferation of allogeneic T lymphocytes.\ Conclusion\ In serum\|free culture,a numbers of dendritic cells was obtained from the human peripheral blood.\;