一种体外扩增NK细胞的新方法

A Novel Method for Expanding NK Cells in Vitro

摘要: 目的：验证一种新型、尚未报道的体外扩增原代NK细胞的方法，同时检测其杀伤活性。方法：采集6位健康志愿者的静脉血共6份，分离外周血单个核细胞。自然杀伤(NK)细胞诱导培养试剂培养14天。利用流式细胞术检测NK细胞所占比例，将K562、THP-1细胞作为靶细胞并用CFSE标记，以效靶比2:1加入扩增后的NK细胞，利用流式细胞术检测靶细胞凋亡。结果：通过14天的细胞培养，利用自然杀伤(NK)细胞诱导培养试剂培养的NK细胞扩增约300倍。NK细胞纯度达50%左右。体外杀伤实验结果显示，扩增后的NK细胞与靶细胞共培养4 h杀伤能力可达40%以上。结论：此种方法扩增的NK细胞其纯度较高，但是因为缺乏其余免疫细胞提供的支撑作用，所以最终NK细胞的扩增倍数并不理想，但其操作简单，花费低，安全性高，可靠性强。

Abstract: Objective: To verify to expand human NK cells ex vivo by using a novel and unreported method and to detect its killing activity. Methods: Mononuclear cells were isolated from 6 samples of peripheral blood from six healthy donors. Natural killer cell induction culture kit was used to culture NK cells from each sample separately for 14 days. The proportion of NK cells was detected by flow cytometry. The K562 and THP-1 cells were used as the targeting cells and the expanded NK cells were added at an effect target ratio of 2:1, and the apoptosis of AML cells was detected by flow cytometry. Results: After 14 days of cell culture, the number of NK cells obtained by natural killer cell induction culture kit for NK cells expanded by 300 times and the ratio of NK cells was about 50.00%. The results of cytotoxicity experiment in vitro showed that the killing ability of NK cells cultured could reach more than 40% when co-cultured with target cells for 4 h. Conclusion: New NK cell culture method using natural killer cell induction culture kit can expand NK cells efficiently and the operation is simple, low cost, high safety and reliability.