摘要
目的:研究组成黄原胶的单糖组分对细胞因子诱导杀伤细胞(CIK)体外扩增的影响。方法:采用正交实验确定最优的单糖组合和添加剂量,并在无血清培养体系中采用细胞培养袋静态培养验证所获得的最优单糖组合对CIK细胞体外扩增的影响。采用流式细胞术检测培养物中各类效应细胞的比例以及效应细胞中表达颗粒酶B和穿孔素的细胞比例;采用CCK-8法检测CIK细胞对K562细胞的杀伤活性。结果:采用添加2.20 mmol/L甘露糖和0.10 mmol/L葡萄糖醛酸的无血清培养基培养单个核细胞时,第14天的总细胞扩增倍数为29.38±5.20,显著高于对照组的18.36±4.03(P<0.05),且效应细胞CD3+、CD3+CD56+、CD3+CD8+细胞的扩增倍数以及CIK细胞中表达颗粒酶B和穿孔素的细胞比例均显著增加(P<0.05),同时扩增后CIK细胞对K562细胞的杀伤活性从(17.93±2.65)%增至(29.03±4.34)%(P<0.05)。结论:甘露糖和葡萄糖醛酸的组合应用可促进CIK细胞体外扩增并提高所获得的CIK细胞的肿瘤杀伤能力。
Objective: To investigate the effect of monosaccharide component constituting xanthan gum on the in vitro expansion of cytokine-induced killer (CIK). Methods: The optimal monosaccharide combination and additive amount was determined based on the orthogonal test.The effect of the optimal monosaccharide combination obtained on the in vitro expansion of CIK cells was verified by static culture in a serum-free culture system in the cellbag.The percentage of effector cells in culture and the percentage of Granzyme B + and Perforin + cells in effector cells were detected by flow cytometry.The killing activity of CIK cells against K562 cells was detected by CCK-8 assay. Results: When mononuclear cells were cultured in serum-free medium supplemented with 2.20 mmol/L mannose and 0.10 mmol/L glucuronic acid,the expansion fold of total cells was up to 29.38±5.20 at day 14,which was significantly higher than that of the control group (18.36±4.03)( P <0.05).The expansion folds of effector cells CD3 +,CD3 +CD56 +,CD3 +CD8 +cells and the percentage of Perforin + cells and Granzyme B + cells in CIK cells were significantly promoted ( P <0.05).The killing activity of expanded CIK cells against K562 cells increased from (17.93±2.65)% to (29.03±4.34)%( P <0.05). Conclusion: The combined application of mannose and glucuronic acid can promote in vitro expansion of CIK cells and enhance the tumor killing ability of the obtained CIK cells.
作者
卢意
蔡海波
谭文松
LU Yi;CAI Hai-Bo;TAN Wen-Song(East China University of Science and Technology,State Key Laboratory of Bioreactor Engineering,Shanghai 200237,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2019年第15期1865-1870,共6页
Chinese Journal of Immunology