不同培养基与血清的培养体系对细胞因子诱导杀伤细胞增殖和功能的影响

发布时间：2018-05-03 23:18

本文选题：细胞因子诱导的杀伤细胞 + 培养基　；参考：《中国肿瘤生物治疗杂志》2014年06期

【摘要】：目的:探讨不同细胞培养基及血清的培养体系对细胞因子诱导杀伤(cytokine-induced killer,CIK)细胞的增殖和功能的影响。方法:采集6例肺癌患者外周血,分离获得单个核细胞,按照不同血清与培养基分为8组:GT-T551培养基+患者自体血清组、GT-T551培养基+健康人血清组、GT-T551培养基+FBS组、GT-T551培养基组、RPMI 1640培养基+患者自体血清组、RPMI 1640培养基+健康人血清组、RPMI 1640培养基+FBS组及RPMI 1640培养基组。采用CFSE染色法检测细胞增殖能力,流式细胞术检测CIK细胞CD3+CD8+T细胞及CD3+CD4+T细胞颗粒酶B、IFN-γ、穿孔素的分泌情况及以白血病NB4、K562细胞为靶细胞时CD3+CD56+T细胞、CD3+CD4+T细胞及CD3+CD8+T细胞表面CD107a的表达。结果:GTT551培养基加入患者自体血清组CIK细胞的增殖指数显著高于其他各组(P0.05)。GT-T551培养基或RPMI 1640培养基中加入患者自体血清组CD4+T细胞颗粒酶B的分泌水平均显著高于加健康人血清组[(22.85±3.50)%vs(13.28±1.75)%,(22.57±3.45)%vs(15.37±4.08)%,均P0.01],而且GT-T551+患者自体血清组CD8+T细胞分泌IFN-γ的能力显著高于加健康人血清组(P0.05)。以白血病细胞系NB4和K562细胞作为靶细胞时,检测CD3+CD56+NKT细胞表面CD107a的表达,GT-T551培养基中加入患者自体血清组优于加健康人血清组[(7.10±1.94)%vs(2.73±0.79)%,(8.00±1.82)%vs(3.03±0.78)%,P0.01],同时优于RPMI1640+患者自体血清组[(4.45±1.96)%、(3.30±1.47)%,P0.01]。结论:GT-T551培养基加患者自体血清的培养体系更有利于CIK细胞的增殖、细胞因子分泌及发挥杀伤功能,可推荐作为CIK细胞最佳培养体系。
[Abstract]:Aim: to investigate the effects of different cell culture medium and serum on the proliferation and function of cytokine-induced killer CIK cells. Methods: mononuclear cells were isolated from peripheral blood of 6 patients with lung cancer. According to different serum and medium, 8 groups were divided into 8 groups: GT-T551, patient's autoserum group, healthy human serum group, FBS medium, GT-T551 medium, patient's autoserum group, RPMI1640 medium, healthy medium. Human serum group: RPMI#number0# medium FBS group and RPMI 1640 medium group. Cell proliferation was detected by CFSE staining. Flow cytometry was used to detect the secretion of CD3 CD8 T cells and CD3 CD4 T cells granzyme Bn- 纬, perforin, and the expression of CD107a on CD3 CD56 T cells, CD3 CD4 T cells and CD3 CD8 T cells. Results the proliferative index of CIK cells in the patients with autologous serum was significantly higher than that in the other groups (P 0.05N. GT-T551 or RPMI 1640). The secretory levels of granzyme B of CD4 T cells were significantly higher in the patients with autologous serum addition than those in the other groups. The ability of CD8 T cells to secrete IFN- 纬 in autologous serum of patients with GT-T551 was significantly higher than that in the serum of healthy persons (22.85 卤1.75 卤22.57 卤3.45)%vs(15.37 卤4.08, P0.01), and the ability of secreting IFN- 纬 by CD8 T cells in autologous serum of GT-T551 patients was significantly higher than that in the control group (P 0.05). When the leukemic cell lines NB4 and K562 cells were used as target cells, the expression of CD107a on the surface of CD3 CD56 NKT cells was detected in GT-T551 medium. The addition of autologous serum from patients was superior to that of healthy persons [7.10 卤1.94)%vs(2.73 卤0.79g + 0.78P 0.01], and it was also better than that of RPMI1640 patients [4.45 卤1.960.30 卤1.47P 0.01]. Conclusion the cell proliferation, cytokine secretion and killing function of CIK cells are more favorable to the culture system of the cell culture medium of: GT-T551 and patient's autologous serum, and can be recommended as the best culture system for CIK cells.
【作者单位】：郑州大学第一附属医院生物细胞治疗中心;郑州大学第一附属医院肿瘤科;郑州大学生命科学院;郑州大学第一附属医院消化内科;河南省高等学校临床医学重点学科开放实验室;
【基金】：国家自然科学基金资助项目(No.81171986,No.81271815) 卫生部科研攻关基金资助(No.20110110001) 河南省科技厅基础与前沿技术研究基金资助(No.112300410153,No.122300410155) 河南省科技厅科技创新人才计划资助(No.124200510006) 郑州大学第一附属医院院内创新团队基金资助~~
【分类号】：R329.2

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