改良RPMI1640培养基对CIK细胞增殖的影响

目的探讨改良RPMI1640培养基对CIK细胞体外增殖的影响,为临床治疗提供质量好、数量多的CIK细胞。方法用淋巴细胞分离液提取淋巴细胞,分别采用传统RPMI1640培养基和改良RPMI1640培养基培养,于培养当天及3、6、9、12、15、18d,采用台盼蓝拒染法计数细胞,检测细胞的增殖水平;流式细胞术分析细胞表型;MTT法检测CIK细胞的体外细胞毒活性。结果培养至第12天后,改良培养基培养条件下,CIK细胞的增殖倍数明显高于传统培养基培养的CIK细胞(P<0.05);CD3+CD56+细胞比例达33%以上;培养15d,对BGC-823和SPC-A-1细胞的细胞毒活性均高于传统培养基。结论改良RP-MI1640培养基较传统RPMI1640培养基可更好地促进CIK细胞增殖,为其临床应用提供了试验数据。

Effect of Modified RPMI1640 Medium on Proliferation of CIK Cells

Objective To investigate the effect of modified RPMI1640 medium on the in vitro proliferation of CIK cells and provide CIK cells at high quality and quantity for clinical therapy. Methods CIK cells were isolated by using Ficoll lymphocyte separating medium, then cultured in traditional and modified RPMI1640 media respectively. On days 0, 3, 6, 9, 12, 15 and 18 after culture, the cells were analyzed for proliferation level by counting with trypan blue exclusion staining, for phenotype by flow cytometry, and for in vitro cytotoxic activity by MTT method. Results The proliferation fold of CIK cells on day 12 after culture in modified RPMI1640 medium was significantly higher than that in traditional medium (P < 0. 05), while the percentage of CD3+ CD56+ cells was more than 33%. Both the cytotoxic activities of CIK cells on day 15 after culture in modified medium to BGC-823 and SPC-A-1 cells were higher than those in traditional medium. Conclusion Compared with traditional RPMI1640 medium, the modified RPMI1640 medium promoted the proliferation of CIK cells effectively, which provided an experimental basis for clinical application of CIK cells.