体外扩增的自然杀伤细胞特点及其杀伤活性研究  

The study on the characteristics and the anti-tumor effects of the natural killer cells expanded in vitro

作　　者：陈稳 毛菊 杨雯雯 胡世莲 程民[1] Chen Wen,Mao Ju,Yang Wenwen,Hu Shilian,Cheng Min(The First Affiliated Hospital of University of Science and Technology of China,Gerontology Institute of Anhui Province,Anhui Provincial Key Laboratory of Tumor Immunotherapy and Nutrition Therapy, Hefei 230001, China)

机构地区：[1]中国科学技术大学附属第一医院(安徽省立医院)安徽省老年医学研究所肿瘤免疫与营养治疗安徽省重点实验室,合肥230001

出　　处：《中国临床保健杂志》2018年第4期531-534,共4页Chinese Journal of Clinical Healthcare

基　　金：安徽省科技攻关项目(1501041142);肿瘤免疫与营养治疗安徽省重点实验室绩效补助(2017070503B041)

摘　　要：目的评价体外扩增的自然杀伤细胞(NK)表型、扩增倍数、杀伤活性以及安全性。方法利用NK细胞体外扩增试剂盒对外周血单个核细胞进行刺激,特异性扩增NK细胞,利用流式细胞术、杀伤实验等检测扩增后NK细胞的特点及抗肿瘤活性;并检测NK细胞外源性因子污染情况等。结果体外培养16~21 d后,细胞扩增500倍以上,总数可达1×1010以上,其中CD3-CD56+NK细胞占90%以上,扩增后的NK细胞对Ho8910、A549、K562、Hep G2及SK-MES-1细胞均有较高的杀伤活性。同时,扩增后NK细胞未检测到细菌、病毒、支原体等外源性病原体污染。结论 NK细胞具有较高的体外杀瘤活性且无外源性因子污染。Objective To evaluate the phenotype,expansion fold,cytotoxic activity and safety of the natural killer cells（ NK cells） expanded in vitro. Methods NK cells reagent kit was used to specifically induce the proliferation of NK cells from the peripheral blood mononuclear cells（ PBMCs）. Flow cytometry assay（ FCM） and in vitro cytotoxicity assay were used to detect the phenotype and the anti-tumor activity of the expanded NK cells. Additionally,the contamination of the exogenous factors of NK cells were also detected. Results After 16-21 days in vitro culture,the PBMC expanded over 500 times to a total number more than 1 × 1010,in which more than 90% of the cells are CD3-CD56＋NK cells. The expanded NK cells displayed high cytotoxicity against Ho8910,A549,K562,Hep G2 and SK-MES-1 cells in vitro. Additionally,no contamination of bacteria,fungus,mycoplasma,or virus was detected in the expanded NK cells. Conclusion The expanded NK cells have high antitumor activity without any toxic side effects in vitro.

关 键 词：肿瘤 杀伤细胞 天然 体外研究 免疫活性 

分 类 号：R73-36[医药卫生—肿瘤]