基于OptiPrep的循环肿瘤细胞富集体系研究  

Study on OptiPrep-based Enrichment System of Circulating Tumor Cell

作　　者：郑罗凝 徐增辉[2] 王真[2] 钱其军[1,2] 

机构地区：[1]浙江理工大学生命科学学院,杭州310018 [2]第二军医大学东方肝胆外科医院,上海200438

出　　处：《浙江理工大学学报（自然科学版）》2015年第5期699-705,共7页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金：国家自然科学基金项目(81071850;81301307)

摘　　要：建立一种简易高效的外周血循环肿瘤细胞(circulating tumor cell,CTC)富集方法。配比不同浓度的OptiPrep溶液梯度离心分离外周血各细胞成分,比较OptiPrep液与现行Ficoll液富集肿瘤细胞的效率、回收率、活性率。以此系统富集肿瘤患者血样中的CTC,结合免疫磁珠筛选、染色体原位荧光杂交和表面抗原免疫荧光技术精确鉴定CTC细胞。结果显示OptiPrep液以1.079g/mL的密度浓度在1 900r/min离心20min为最优分离条件。32例临床肿瘤患者血样的CTC检测实验中,均发现有循环肿瘤细胞的存在。结果表明基于OptiPrep液结合免疫磁珠筛选能有效富集外周血循环肿瘤细胞,为实验室常规开展CTC的计数、培养等研究奠定基础。This study aimed to establish a simple and efficient enriching method of CTC（circulating tumor cell,CTC）in the peripheral blood.OptiPrep solutions with different concentration was prepared to separate different types of cells in peripheral blood,and the efficiency of enrichment,recovery,and cell viability of OptiPrep solution and Ficoll solution were compared.This system was used to enrich CTC in blood samples of cancer patients.CTC cells were identified in combination of immunomagnetic beads,chromosome fluorescence in situ hybridization and surface antigen immunofluorescence technique.The results show that the optimal separation condition is as follows：OptiPrep density 1.079g/mL;the centrifugation speed and time 1 900r/min,20 min.During CTC examination of blood samples of 32 enrolled cancer patients,it is found that there exist circulating tumor cells.This study demonstrats that OptiPrep-based enrichment system combined with immunomagnetic beads could effectively capture circulating tumor cells in peripheral blood,which flays a foundation for CTC count,culture and other researches in laboratory.

关 键 词：OptiPrep分离介质 密度梯度离心法 循环肿瘤细胞 免疫磁珠 

分 类 号：R73-37[医药卫生—肿瘤]