Luzyanina Tatyana, Cupovic Jovana, Ludewig Burkhard, Bocharov Gennady
Institute of Mathematical Problems in Biology, RAS, Pushchino, Russia,
J Math Biol. 2014 Dec;69(6-7):1547-83. doi: 10.1007/s00285-013-0741-z. Epub 2013 Dec 13.
Since their invention in 1994, fluorescent dyes such as carboxyfluorescein diacetate succinimidyl ester (CFSE) are used for cell proliferation analysis in flow cytometry. Importantly, the interpretation of such assays relies on the assumption that the label is divided equally between the daughter cells upon cell division. However, recent experimental studies indicate that division of cells is not perfectly symmetric and there is unequal distribution of protein between sister cell pairs. The uneven partition of protein or mass to daughter cells can lead to an overlap in the generations of CFSE-labelled cells with straightforward consequences for the resolution of individual generations. Numerous mathematical models developed so far for the analysis of CFSE proliferation assay incorporate the premise that the CFSE fluorescence intensity is halved in the two daughter cells. Here, we propose a novel modelling approach for the analysis of the CFSE cell proliferation assays which are characterized by poorly resolved peaks of cell generations in flow cytometric histograms. We formulate a mathematical model in the form of a system of delay hyperbolic partial differential equations which provides a good agreement with the CFSE histograms time-series data and allows an analytical treatment. The model is a further generalization of the recently proposed class of division- and label-structured models as it considers an asymmetric cell division. In addition, the basic structure of the cell cycle, i.e. the resting and cycling cell compartments, is taken into account. The model is used to estimate fundamental parameters such as activation rate, duration of the cell cycle, apoptosis rate, CFSE decay rate and asymmetry factor in cell division of monoclonal T cells during cognate interaction with dendritic cells.
自1994年发明以来,诸如羧基荧光素二乙酸琥珀酰亚胺酯(CFSE)之类的荧光染料被用于流式细胞术中的细胞增殖分析。重要的是,此类检测的解读依赖于这样一种假设,即细胞分裂时标记物在子代细胞之间平均分配。然而,最近的实验研究表明,细胞分裂并非完全对称,姐妹细胞对之间的蛋白质分布不均。蛋白质或质量向子代细胞的不均匀分配会导致CFSE标记细胞的代际重叠,这对分辨各个代际会产生直接影响。到目前为止,为分析CFSE增殖检测而开发的众多数学模型都包含CFSE荧光强度在两个子代细胞中减半的前提。在此,我们提出一种用于分析CFSE细胞增殖检测的新颖建模方法,这类检测在流式细胞术直方图中细胞代际峰分辨率较差。我们构建了一个以延迟双曲型偏微分方程组形式的数学模型,该模型与CFSE直方图时间序列数据吻合良好,并允许进行解析处理。该模型是对最近提出的一类分裂和标记结构模型的进一步推广,因为它考虑了不对称细胞分裂。此外,还考虑了细胞周期的基本结构,即静止和循环细胞区室。该模型用于估计诸如激活率、细胞周期持续时间、凋亡率、CFSE衰减率以及单克隆T细胞在与树突状细胞同源相互作用期间细胞分裂的不对称因子等基本参数。