Generation of thymocyte subpopulations in organ culture: correlated analysis of Lyt-2 phenotype and cell cycle status by flow microfluorometry.
Generation of thymocyte subpopulations in organ culture: correlated analysis of Lyt-2 phenotype and cell cycle status by flow microfluorometry.
复制标题
器官培养中胸腺细胞亚群的产生：通过流式显微荧光测定法对 Lyt-2 表型和细胞周期状态进行相关分析。
作者：
R. Sékaly;R. Ceredig;H. Macdonald
我们已经研究了细胞周期动力学和增殖活性的Lyt-2定义的亚群在器官培养的胸腺雏形。在妊娠13天取出并培养4 - 7天的胎儿胸腺与溴脱氧尿苷孵育，然后用Hoechst 33342染色以定量细胞周期的持续时间和周期细胞的数量。细胞周期的总持续时间为30 hr. The器官培养细胞根据其Lyt-2表型和其前向光散射的分选，然后Hoechst荧光和碘化丙啶分析，表明小Lyt-2+细胞，本身是noncycling来自通过有丝分裂从另一个细胞群体。Hoechst-BrdU替代技术与单克隆抗体和器官培养相结合，将为胸腺谱系通路的研究提供有力的工具。
We have investigated the cell cycle kinetics and the proliferative activity of Lyt-2-defined subpopulations in organ-cultured thymic rudiments. Fetal thymuses that were removed at 13 days of gestation and cultured for 4 to 7 days were incubated with bromodeoxyuridine and were then stained with Hoechst 33342 to quantitate the duration of the cell cycle and the number of cycling cells. The total duration of the cell cycle was 30 hr. The sorting of organ culture cells according to their Lyt-2 phenotype and their forward light scatter, followed by Hoechst fluorescence and propidium iodide analysis, indicated that small Lyt-2+ cells that are themselves noncycling were derived through mitosis from another cell population. The combination of the Hoechst-BrdU substitution technique with monoclonal antibodies and the organ culture should provide a powerful tool for the study of lineage pathways in the thymus.