Percoll及Ficoll分离纯化肝癌移植瘤肿瘤浸润淋巴细胞比较  

Comparison of Percoll and Ficoll isolated and purified tumor infiltrating lymphocytes in hepatocellular carcinoma

作　　者：孙盼[1] 樊晓晖[1] 张彬[1] 张磊[1] 蔡双启[2] 梁莹[1] 赖振屏[1] 高灵茜[1] 宋德志[1] Sun Pan;Fan Xiaohui;Zhang Bin;Zhang Lei;Cai Shuangqi;Liang Ying;Lai Zhenping;Gao Lingqian;Song Dezhi(Department of Microbiology,College of Basic Medical Sciences,Guangxi Medical University 530021;Institute of Respiratory Diseases,the First Affiliated Hospital of Guangxi Medical University 530021)

机构地区：[1]广西医科大学基础医学院微生物学教研室,530021 [2]广西医科大学第一附属医院呼吸疾病研究所,530021

出　　处：《中国社区医师》2016年第13期7-8,10,共3页Chinese Community Doctors

基　　金：国家自然科学基金项目(81460437);广西自然科学基金青年基金项目(2013GXNSFBA019160);广西科技技术开发项目(桂科合1347004-27);教育部博士导师联合基金项目(20124503110007);广西特聘专家岗位科研项目(2013-2016);广西教育厅科研立项项目(201204LX032);广西卫生厅课题项目;桂卫自筹Z2013045;广西壮族自治区卫生和计划生育委员会科研课题:Z2015500~~

摘　　要：目的:比较Percoll和Ficoll两种淋巴细胞分离液对肿瘤浸润淋巴细胞纯度及细胞增殖的影响。方法:用Percoll及Ficoll两种淋巴细胞分离液分离H22肝癌小鼠皮下移植瘤内淋巴细胞,对细胞首先行锥虫蓝染色后在显微镜下计算细胞总数和存活率,最后用流式细胞仪分析淋巴细胞纯度及分裂、增殖能力。结果:Percoll及Ficoll分离所得TIL总数分别为(3.28±0.78)×107和(2.02±0.76)×107;Percoll分离所得第3、5、7天的TIL增殖率分别为(22.8±2.31)%、(39.1±1.94)%、(69.85±1.68)%;Ficoll分离所得第3、5、7天的TIL增殖率分别为(13.25±1.68)%、(23.7±5.31)%、(41.5±1.56)%。Percoll分离所得TIL中CD3+T、CD4+T、CD8+T细胞分别为(73.2±6.8)%、(49.1±10.2)%、(22.0±1.4)%;Ficoll分离所得TIL中CD3+T、CD4+T、CD8+T淋巴细胞分别为(47.9±5.0)%、(29.1±8.4)%、(19.7±5.0)%。结论:与Ficoll相比,Percoll分离所得H22肝癌小鼠皮下移植瘤TIL的细胞得率和纯度更高、增殖力较强,将更有利于后续研究。Objective:To compare the effects of the two kinds of Percoll and Ficoll lymphocytes on the purity and cell proliferationof tumor infiltrating lymphocytes.Method:H22 mice were isolated subcutaneous transplantation tumor lymphocyte by Percoll andFicoll two kinds of lymphocytes.The cell count and survival rate were calculated under microscope after the cells were firststained with a cell line,and the purity and proliferation ability of lymphocytes were analyzed by flow cytometry.Results:The totalnumber of Percoll and Ficoll isolated from TIL respectively(3.28±0.78)×107 and(2.02±0.76)×107;the TIL proliferation rate ofthe 3d,5d,7d through Percoll separating income were(22.8 ± 2.31% ),(39.1 ± 1.94% ),(69.85 ± 1.68)% respectively;through theFicoll separating income were (13.25±1.68)%,(23.7±5.31)% and(41.5±1.56)% respectively.The CD3+T,CD4+T,CD8+T cells inTIL through Percoll separating income were(73.2±6.8%),(49.1±10.2%),(22.0±1.4)%;the CD3+ T,CD4+ T,CD8+ T cells in TILthrough Ficoll separating income were(47.9 ± 5)% and(29.1 ± 8.4)% and(19.7 ± 5)% .Conclusion:Compared with Ficoll,theseparation of Percoll H22 mice transplanted TIL cell yield and purity is higher,stronger proliferation ability,which will be moreconducive to the follow-up study.

关 键 词：肿瘤浸润淋巴细胞 淋巴细胞分离液 密度梯度分离 肝癌移植瘤 

分 类 号：R735.7[医药卫生—肿瘤]