外周血单个核细胞与HepG2.215细胞共培养体系的研究

目的建立外周血单个核细胞与HepG2.215细胞共培养体系,探讨不同培养基及效靶比对共培养体系的影响。方法分离正常人外周血单个核细胞,加入植物血凝素（PHA）,置于不同的培养基（DMEM、RPMI1640）中进行培养,采用不同效靶比（5∶1、10∶1、20∶1、40∶1）构建PBMCs与HepG2.215细胞共培养体系。用倒置显微镜观察细胞的形态及生长情况,台盼蓝拒染法检测PBMCs与HepG2.215细胞的细胞活力,cck-8法检测HepG2.215细胞的增殖活性。结果 DMEM培养基培养的HepG2.215细胞的细胞活力比RPMI1640培养基培养的细胞高;而两种培养基培养的PBMCs的细胞活力无明显变化。共培养条件下,PBMCs对HepG2.215细胞增殖活性的抑制作用随效靶比的不同而有所差别,效靶比为20∶1时抑制作用最强。结论在PBMCs与HepG2.215细胞共培养体系中,细胞培养基和效靶比对HepG2.215细胞的生长有影响。

Study of peripheral blood mononuclear cells and HepG2.215 cells co-culture system

Objective To establish the co-culture system of peripheral blood mononuclear cells and HepG2.215 cells,and explore the effects of different culture medium and the target ratio on the impact of co-culture system.Methods PBMCs separated from normal human blood were cultured with PHA in different mediums（DMEM,RPMI1640）.The co-culture system of PBMCs and HepG2.215 cells was established in different proportion（5∶1,10∶1,20∶1,40∶1）.Morphology and growth condition of cells were observed by inverted microscope.The activity of PBMCs and HepG2.215 cells was evaluated by Trypan blue staining.The proliferation of HepG2.215 cells was measured by cck-8.Results The activity of HepG2.215 cells cultured with DMEM was higher than that of cells cultured with RPMI 1640（P0.05）.The activity of PBMCs had no significant change.The inhibition of PBMCs on HepG2.215 cells changed with the ratio of effector cells and target cells in co-culture condition,and shoed the strongest inhibition when the target ratio was 20∶1.Conclusion In the PBMCs and HepG2.215 cells co-culture system,proliferation of HepG2.215 cells is effected by culture medium and the target ratio.