一种高倍扩增细胞因子诱导杀伤细胞方法  

A high-efficiency system of cytokine induced killer(CIK) cell expansion

作　　者：段彦龙[1] 许凝[2] 杨桦[1] 安利佳[1] 

机构地区：[1]大连理工大学环境与生命学院,辽宁大连116024 [2]大连大学附属新华医院胸外科,辽宁大连116021

出　　处：《大连理工大学学报》2008年第3期334-338,共5页Journal of Dalian University of Technology

基　　金：大连市科学技术基金资助项目(2004134SF119)

摘　　要：CD3单克隆抗体分别以包被和悬浮的方式刺激人外周血单个核细胞,并以不同的基本培养基(RPMI-1640、IMDM、DMEM/F-12(1∶1)和M199)以及抗氧化剂(2-巯基乙醇和亚硒酸钠)培养细胞因子诱导杀伤(CIK)细胞,分别测定细胞的扩增、表型(CD3CD56和CD25)和非MHC-限制性毒性(NK毒性和LAK毒性).结果显示高浓度CD3单克隆抗体包被刺激方式的细胞集落和细胞扩增倍数大于悬浮刺激方式.基本培养基DMEM/F-12(1∶1)的扩增倍数优于其他3种基本培养基.抗氧化剂2-巯基乙醇和亚硒酸钠均有促进CD25抗原形成的作用,亚硒酸钠能提高CIK细胞的毒性,而2-巯基乙醇对于细胞的扩增有促进作用.建立了一种高倍扩增CIK细胞的方法,20 d细胞平均扩增倍数可以达到千倍以上.Activation of peripheral blood mononuclear cells （PBMCs） with different means including immobilized and suspended anti-CD3 monoclonal antibody, cytokine induced killer（CIK） cells were induced by culturing PBMCs in the basal media （RPMI-1640, IMDM, DMEM/F-12 （1 ： 1） and M199） and antioxidant （2-mercaptoethanol and sodium selenite） in vitro, the expansion rates,surface markers （CD3CD56 and CD25） and non-MHC restriction cytotoxicities （NK cytotoxicity and LAK cytotoxicity） were evaluated. The results show that colonies and the expansion rates of the cells activated by immobilized anti-CD3 monoclonal antibody are more than those of the suspended antibody. The mean expansion rates of the basal medium DMEM/F-12 （1 ： 1） is the highest in the four media. 2-mercaptoethanol and sodium selenite as antioxidants in the basal medium DMEM/F-12 （1 ： 1） can increase CD25 surface markers, 2-mercaptoethanol can increase the expansion rates, and sodium selenite can increase the cytotoxicities. In conclusion, a high-efficiency system of CIK cells expansion is built by this system, which can approximately achieve three thousand-fold expansions of the total cells within 20 days in vitro.

关 键 词：细胞因子诱导杀伤(CIK)细胞 细胞扩增 生物反应器 

分 类 号：R318[医药卫生—生物医学工程]