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目的 研究F344大鼠树突状细胞(DC)的分离与培养方法。方法 取大鼠四肢骨骨髓细胞悬液,经梯度离心法得到DC ,加入白细胞介素(IL) - 4、粒细胞巨噬细胞集落刺激因子(GM CSF) ,全RPMI 16 4 0培养,经倒置显微镜、电镜及流式细胞仪鉴定细胞纯度及成熟度。结果 细胞培养的第8天,经倒置显微镜、电镜观察DC出现典型形态;流式细胞仪检测第4天OX6 2阳性率为8 .92 %、主要组织相容性复合物(MHC) Ⅱ为2 0 . 90 %、CD86为16 98% ;第8天OX6 2阳性率为5 8. 0 7%、MHC Ⅱ为6 0 . 4 9%、CD86为6 2 . 94 % ;第15天OX6 2阳性率为84 . 6 8%、MHC Ⅱ为88 .0 3%、CD86为6 2 . 80 %。结论 用该方法得到的DC纯度可达80 %以上,为进一步研究其在肿瘤研究中的应用打下基础。
Objective To study the isolation and culture of dendritic cells (DCs) from F344 rats. Methods The myeloid bone marrow cell suspension was obtained from the extremities of rats. The DCs were obtained by gradient centrifugation. After addition of interleukin - 4 (GM - CSF) and GM - CSF, Microscopy, electron microscopy and flow cytometry to identify cell purity and maturity. Results On the 8th day of cell culture, the typical morphology of DCs was observed by inverted microscope and electron microscope. The positive rate of OX6 2 was 8.92% on flow cytometry on day 4, and the major histocompatibility complex (MHC) Ⅱ was 2 0 90% and CD 98 16 98%. The positive rate of OX6 2 on the 8th day was 58.07%, the MHC Ⅱ 60.99% and the CD86 6 694.9% The rate was 84.68%, MHC Ⅱ was 88.0%, CD86 was 62.8%. Conclusion The purity of DC obtained by this method can reach more than 80%, which lays the foundation for further studying its application in tumor research.