利用活细胞工作站构建巨噬细胞吞噬子宫内膜癌细胞的体外模型

In vitro phagocytosis model of endometrial cancer cells by macrophages via live cell application

目的利用活细胞工作站观察巨噬细胞吞噬子宫内膜癌细胞的过程及流式细胞术检测巨噬细胞吞噬肿瘤细胞对T细胞的活化。方法将Ishikawa细胞与THP-1诱导的巨噬细胞分别用CFSE荧光探针和CD11b标记,混合接种于成像皿,实时记录120 min内巨噬细胞对Ishikawa细胞的吞噬,流式细胞术检测巨噬细胞与Ishikawa细胞共培养后对T细胞活化的影响。结果共培养的2种细胞相互接触后Ishikawa细胞的绿色荧光被巨噬细胞摄取,且巨噬细胞能连续吞噬多个Ishikawa细胞。吞噬了Ishikawa细胞的巨噬细胞能诱导细胞毒性T细胞分化。结论成功利用活细胞工作站构建巨噬细胞吞噬肿瘤细胞的体外模型,为检测巨噬细胞及T细胞对肿瘤细胞双重杀伤作用提供可行的实验方法。

Objective The live cell application was used to observe the process of phagocytosis of endometrial cancer cells by macrophages,and flow cytometry was used to detect the effects of macrophages engulfing tumor cells on activation of cytotoxic T cell.Methods Ishikawa cells and THP1-induced macrophages were labeled with CFSE fluorescent probe and CD11b respectively,and then mixed and seeded on a glass imaging dish.The live cell application was performed to record the phagocytosis of Ishikawa cells by macrophages within 120 minutes.Flow cytometry was used to detect the effect of macrophages engulfing tumor cells on activation of cytotoxic T cell.Results The green fluorescence of Ishikawa cells was taken up by macrophages after the co-cultured two types of cells were in contact with each other,and macrophages were able to engulf multiple Ishikawa cells continuously.Macrophages that engulfed Ishikawa cells could induce activation of cytotoxic T cell.Conclusion The live cell application was successfully conducted to construct an in vitro model of phagocytosis of tumor cells by macrophages,which provided a feasible experimental method for detecting the dual killing process of macrophages and T cells on tumor cells.