Schwarting R, Gerdes J, Niehus J, Jaeschke L, Stein H
J Immunol Methods. 1986 Jun 10;90(1):65-70. doi: 10.1016/0022-1759(86)90384-4.
A novel procedure for determining the growth fraction of cell suspensions by flow cytometry is described. This method identifies proliferating cells by binding the monoclonal antibody Ki-67 to a nuclear antigen present in all cells that are in the G1, S, G2, and M phase of the cell cycle, but not in the G0 phase. In a kinetic study of Na cell line U937 using concanavalin A for stimulation of peripheral blood mononuclear cells, a steady increase of Ki-67 positive cells evaluated by flow cytometry was observed. Simultaneously, the [3H]thymidine uptake of the ConA blasts was measured and compared to the expression of Ki-67. A linear correlation between the percentage of Ki-67 positive cells and the log transformed counts per minute was demonstrated, and staining with Ki-67 detected cell proliferation with the same sensitivity as 3H-TdR uptake. In addition, it was possible to stain Ki-67-labelled cells with a second cell marker if a second fluorescent dye coupled to an antibody was used. This provided the opportunity to define precisely the phenotype of proliferating cells. Conversely, the number of proliferating cells expressing certain preselected surface markers could be easily determined.
本文描述了一种通过流式细胞术测定细胞悬液生长分数的新方法。该方法通过将单克隆抗体Ki-67与细胞周期中G1、S、G2和M期所有细胞中存在的核抗原结合来识别增殖细胞，而G0期细胞中不存在该核抗原。在一项关于Na细胞系U937的动力学研究中，使用伴刀豆球蛋白A刺激外周血单核细胞，通过流式细胞术评估发现Ki-67阳性细胞稳步增加。同时，测量伴刀豆球蛋白A刺激的母细胞的[3H]胸苷摄取，并与Ki-67的表达进行比较。结果表明，Ki-67阳性细胞百分比与每分钟对数转换计数之间呈线性相关，并且用Ki-67染色检测细胞增殖的灵敏度与3H-TdR摄取相同。此外，如果使用与抗体偶联的第二种荧光染料，则可以用第二种细胞标记物对Ki-67标记的细胞进行染色。这为精确确定增殖细胞的表型提供了机会。相反，表达某些预选表面标志物的增殖细胞数量也可以很容易地确定。
