【摘要】 目的探讨细胞膜染料PKH26在淋巴细胞及其亚群增殖中的应用。方法利用PKH26染色、荧光抗体标记和流式细胞术,检测淋巴细胞及其亚群在多克隆刺激剂刺激作用下荧光强度的变化,应用相关软件分析其增殖情况,并与传统MTT方法进行比较。结果经不同浓度的多克隆刺激剂刀豆蛋白A(ConA)刺激72h后,两种方法均能检测淋巴细胞增殖情况,其增殖强度与刺激剂浓度呈正相关(r=0.999)。经同种细胞刺激6d后,结合CD8、CD3抗体染色,经ModFit软件拟合后,PKH26能显示的T细胞亚群与T淋巴细胞明显不同的细胞增殖动力。结论PKH26染色结合荧光抗体标记和流式细胞术,是分析淋巴细胞及其亚群增殖动力的有力工具。更多还原

【Abstract】 Objective To explore the application of PKH26 in the proliferation of lymphocytes and their subsets. Methods PKH26 staining, fluorescence antibody labeling, flow cytometry and related software were used to detect the fluorescence intensity and analyze the proliferation kinetics of lymphocytes and their subsets after stimulation with polyclonal stimulators or allogeneic cells. Results Lymphocytes divided after stimulation of ConA for 72 h. The proliferation index of lymphocytes using PKH26 gave results consistent with a parallel MTT assay (r= 0.999). The proliferative kinetics obtained by use of the “ModFit” software showed that the proliferative kinetics of the subsets was different from the total T cell primed by allogeneic cell. Conclusion PKH26 staining combined with fluorescent antibody labeling and flow cytometry were powerful tools for analysis of proliferation of lymphocytes and their subsets.更多还原