接种密度对C57BL/6小鼠CIK细胞增殖、分化及杀瘤作用的影响  

Effects of Different Inoculation Density on the Proliferation, Differentiation and Anti-Tumor Capability of CIK Cells in C57BL/6 Mice

作　　者：李光宗[1] 曾喜欢 刘英富[3] 霍景瑞[3] 郁硕 张益 侯世科[1] 陈小义[3] 陈锋[1] 

机构地区：[1]武警后勤学院附属医院救援医学研究所,天津300162 [2]锦州医科大学,辽宁锦州121001 [3]武警后勤学院细胞生物与医学遗传学教研室,天津300309

出　　处：《生物技术通讯》2017年第3期342-346,共5页Letters in Biotechnology

基　　金：天津市科技计划(14ZCDZSY00033);全军重点实验室开放基金(JY1402);武警后勤学院附属医院科研平台开放基金(WYFK-FM201602;WYKFZ201603)

摘　　要：目的:探讨不同接种密度对C57BL/6小鼠细胞因子诱导的杀伤细胞(CIK细胞)增殖分化及杀瘤作用的影响,进一步优化CIK细胞培养方法。方法:按照1×10~6/mL(A组)、4×10~6/mL(B组)、8×10~6/mL(C组)、12×10~6/mL(D组)4种接种密度培养细胞,加入必要的细胞因子,14 d后收获细胞,通过流式细胞术、CCK-8法对细胞增殖、分化、杀瘤作用进行分析。结果:培养过程中,C组细胞形态及增殖能力优于A、B组,在14 d时收获细胞并对其进行检测时发现,C组CD3^+/NK1.1^+细胞所占比例明显高于A、B组,杀瘤活性也优于A、B组;D组细胞密度过大,在7 d细胞进入快速增殖期后出现大面积死亡。结论:适当提高C57BL/6小鼠CIK细胞的接种密度利于细胞增殖、分化及杀瘤作用的形成,选择8×10~6/mL的接种密度是较为合适的。Objective： To investigate the effects of different inoculation densities on proliferation, differentiation and anti-tumor capability of cytokine-induced killer（CIK） cells in C57BL/6 mice, and to further optimize CIK cells culture method.Methods： The cells were cultured in the density of 1×10^6/mL（group A）, 4×10^6/mL（group B）, 8×10^6/mL（group C） and 12×10^6/mL（group D）, and after 14 days, the cells were harvested and analyzed by microscope, flow cytometry and CCK-8 for cell proliferation, differentiation and anti-tumor capability.Results： The cell morphology and proliferation ability of group C were superior to those of group A and B. Whencells were harvested and detected on 14 d, the percentage of CD3^＋/NK1.1^＋ cells in C group was significantly higher than the proportion of group A and B, anti-tumor capability was better than that of group A and B. Cell density of group D cell density was too large, in the 7 d cells into the rapid proliferation phase, the cells appear large area of death.Conclusion： It is appropriate to increase the inoculation density of CIK cells in C57BL/6 mice, which is beneficial to the proliferation, differentiation and anti-tumor capability of C57BL/6 mice, and the inoculation density of 8×10^6/mL is more appropriate.

关 键 词：细胞因子诱导的杀伤细胞 密度 细胞培养 增殖 杀瘤作用 

分 类 号：Q25[生物学—细胞生物学]