非小细胞肺癌EMPE来源树突状细胞的分离培养

The Isolation and Culture of Dendritic Cells Derived from EMPE in Nsclc

目的:建立非小细胞肺癌(non-small cell lung cancer,NSCLC)渗出性恶性胸腔积液(exudativemalignant pleural effusions,EMPE)来源树突状细胞(dendritic cells,DCs)的分离培养方法,观察DCs的形态学及对其进行表型鉴定。方法:用密度梯度离心的方法,从非小细胞肺癌EMPE中分离获得胸腔积液中的单个核细胞,用白细胞介素-4(IL-4)、粒细胞巨噬细胞集落刺激因子(granulocyte macrophage colony-stimulating factor,GM-CSF)、肿瘤坏死因子-α(TNF-α)培养,应用倒置显微镜、光镜观察细胞形态;用流式细胞仪对DCs进行表型鉴定。结果:建立了PEDCs的分离培养方法。培养9 d的DCs高表达人类白细胞抗原-DR(human leucocyte antigen,HLA-DR)、CD83、CD86、CD80。结论:从EMPE中分离的PEDCs体外培养后不仅在形态上成熟,而且高表达特异性表面分子。

Objective：To establish the method of isolation and culture of dendritic cells derived from exuda-tive malignant pleural effusions （EMPE） of non - small cell lung cancer and to make the morphological observa- tion and phenotype analyzation. Methods： Using the method of density gradient centrifugation, we obtained malignant effusion - associated monocytes. Then we cultured DC, s using IL - 4 , GM - CSF and TNF - a. The morphological properties of DCs were observed by inverted microscope and optical microscope. The phenotypes of DCs were analyzed by flow cytometry. Results： The method for isolation and culture of PEDCs . DCs cultured for 9 days expressed highly HLA- DR ,CD83, CD86, CD80. Conclusion： PEDCs isolated from EMPE and cultured in vitro got to maturation in morphology and expressed typital surface phenotypes of mature DCs.