摘要
目的观察脐血来源树突状细胞(DC)与细胞因子诱导的杀伤细胞(CIK)共培养(DC-CIK)对结肠癌细胞HT29的杀伤作用,探讨脐血来源DC-CIK替代外周血来源DC-CIK的可行性。方法取脐血及外周血各50 m L,密度梯度离心法分离单个核细胞,诱导培养7天至DC成熟,将成熟DC和CIK混合培养8天,观察DC-CIK的形态变化;培养4、8、12、16天检测不同来源CIK/DC-CIK的增殖情况,培养前及培养第7天DC表型(CD40+、CD80+、CD86+),培养前CIK及培养第16天DC-CIK CD3+、CD3+CD56+比例。按效应细胞(脐血或外周血DC-CIK)∶靶细胞(HT29细胞)=5∶1、10∶1、20∶1混合,培养24 h,采用CCK-8法检测不同来源DC-CIK对HT29细胞的增殖抑制率,流式细胞仪检测细胞凋亡率。结果脐血及外周血来源DC、DC-CIK形态培养前后无明显差别,CD40+、CD80+、CD86+DC所占比例培养前后比较均无明显统计学差异(P均>0.05)。体外诱导培养第16天,脐血来源CD3+及CD3+CD56+DC-CIK增殖能力显著高于外周血来源DC-CIK,不同效靶比脐血来源DC-CIK对HT29细胞的增殖抑制及促凋亡作用效果均优于外周血来源DC-CIK(P均<0.05)。结论脐血来源DC-CIK可替代外周血来源DC-CIK作为肿瘤免疫治疗的种子细胞。
Objective To investigate the killing effect of co-cultured umbilical cord blood-derived dendritic cells （DC） and cytokine-induced killer cells （CIK） on colon cancer cells HT29, to explore the feasibility of using umbilical cord blood derived DC-CIK to substitute peripheral blood derived DC-CIK.Methods Umbilical cord blood （50mL） and peripheral blood （50 mL） was collected respectively.The mononuclear cells were isolated by density gradient centrifugation under aseptic conditions, and DC cells were induced and cultured until their mature.The matured DC and CIK cells were mixed for 8 d, the morphological changes of DC-CIK were observed by inverted microscope.The proliferation of CIK/DC-CIK from umbilical cord blood and peripheral blood were evaluated by cell count assay on day 4, 8, 12 and 16, and specif-ic markers of CIK/DC-CIK （ CD3 ＋, CD3 ＋CD56 ＋） were identified by flow cytometric immunophenotype analysis on day 0 and 16, the specific markers of DC （CD40, CD80, and CD86） were tested on day 0 and 7.The DC-CIK cells were co-cultured in different concentration ratio with HT29 cells （DC-CIK∶HT29 =5∶1, 10∶1 and 20∶1） for 24 h, and the antitu-mor rate of DC-CIK on HT29 was detected by CCK-8, and the apoptosis rate of HT29 was investigated by flow cytometry. Results There was no significant difference in the cellular morphology between umbilical cord blood-derived and peripher-al blood-derived DC/DC-CIK before and after culturing, as well as in the specific markers of DC （ CD40, CD80, and CD86）.After 16 days culture, the proliferation rate of CD3 ＋, CD3 ＋CD56 ＋cells from umbilical cord blood was increased significantly as compared with that from the peripheral blood.The stronger killing effect and apoptosis-promoting effect on HT29 cells was found in umbilical cord blood-derived DC-CIK as compared with that of the peripheral blood derived DC-CIK （P〈0.05）.Conclusion The umbilical cord blood-derived DC-CIK can replace peripheral blood-derived DC-CIK as＆nbsp;the
出处
《山东医药》
CAS
北大核心
2016年第40期13-16,共4页
Shandong Medical Journal
基金
山东省自然科学基金高校
科研单位联合专项计划(ZR2015HL063)
山东省医药卫生科技发展计划项目(2014WS0300)
关键词
结肠癌
树突状细胞
细胞因子诱导的杀伤细胞
脐血
外周血
colorectal carcinoma
dendritic cells
cytokine-induced killer cells
cord blood
peripheral blood
