摘要
【目的】尝试使用IL-2、IL-7和IL-12的单因子、双因子和三因子组合进行杀伤细胞/自然杀伤细胞(CIK/NK)细胞扩增培养,探讨最优化CIK/NK细胞少因子培养体系的建立。【方法】采用Ficoll-Hypaque法分离脐血单个核细胞并分成6组,每组分别加入IL-2(80ng/mL)、IL-7(40ng/mL)及IL-12(40ng/mL)的单因子、双因子和三因子组合(IL-7组;IL-12组;IL-7+IL-12组;IL-2+IL-7组;IL-2+IL-12组;IL-2+IL-7+IL-12组),每组6个重复孔,共培养21d收获细胞。在细胞培养开始和第21天,实验各组取细胞悬液进行流式细胞仪检测CD3+CD56+CIK细胞及CD3-CD56+NK细胞比例。【结果】单因子培养体系的IL-12组在CIK细胞扩增比例上是各组中最高的(22.96±1.41)%,双因子体系的IL-2+IL-12组的CIK细胞扩增比例也达到(18.58±0.68)%;上述两组的CIK细胞扩增比例已经达到或基本达到使用传统多因子扩增方法所获得的比例。IL-12组的NK细胞扩增比例为(30.23±1.18)%,IL-2+IL-7组的NK细胞扩增比例也达(29.52±0.89)%。【结论】使用IL-12、IL-2+IL-12或IL-2+IL-7组合的少因子培养体系进行CIK/NK细胞的培养扩增是可行的。
[Objective] In order to explore the optimal minor-factor culture system of CIK/NK cells by proliferating CIK/NK cells using single-factor, bi-factor, and tri-factor combinations of IL-2, IL-7, and IL-12. [Methods] Ficoll-Hypaque method was used to separate cord blood mononuclear cells and divide them into 6 groups. Add single-factor, bi-factor, and tri-factor combinations （IL-7; IL-12; IL-7 ＋ IL-12; IL-2 ＋ IL-7; IL-2 ＋ IL-12; IL-2 ＋ IL-7 ＋ IL-12） of IL-2 （80 ng/mL）, IL-7 （40 ng/ mL）, and IL-12 （40 ng/mL） and culture them in total 21 days then harvest the cells. To collect cell suspensions of each group in culture outset and day 21 and to count the proportion of CD3^＋ CD56^＋CIK cells and CD3^- CD56^＋ NK cells with flow cytometry. [Results] The proportion of CIK cells in single-factor culture system of IL-12 was the highest in all groups （30.23 ± 1.18%）. The proportion of CIK cells in bi-factor culture system of IL-2 ＋ IL-12 can raised to 18.58 ± 0.68%. The proportion of CIK cells of the two groups above can reach the level of that using traditional multi-factor culture methods. NK cell proportion in IL-12 was 30.23 ± 1.18%, NK cell proportion in IL-2 ＋ IL-7 can also reach to 29.52 ± 0.89%. [Conclusions] It is adoptable to proliferate CIK/NK cells using minor-factor culture system of IL-12, IL-2 ＋ IL-12, or IL-2 ＋ IL-7.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2009年第4期361-366,共6页
Journal of Sun Yat-Sen University:Medical Sciences
基金
国家自然科学基金(30500469)
广东省科技计划项目(2008B030301066)
广州科学仪器协作共用网基金(2006034)
中山大学北校区学生科研基金(2006031)
