Pereira Túlio Felipe, Levin Gabriel, DeOcesano-Pereira Carlos, Caodaglio Amanda Schiersner, Fujita André, Tonso Aldo, Sogayar Mari Cleide
Cell and Molecular Therapy Center (NUCEL), Department of Internal Medicine, School of Medicine, University of São Paulo, Rua Pangaré, 100, Cidade Universitária, São Paulo, SP, 05360-130, Brazil.
Department of Biochemistry, Chemistry Institute, University of São Paulo, São Paulo, SP, Brazil.
BMC Res Notes. 2020 Feb 4;13(1):57. doi: 10.1186/s13104-020-4914-8.
Cell growth curves constitute one of the primary assays employed to analyze cell proliferation dynamics of in vitro cultured cells under specific culture conditions. From the cell growth curve, it is possible to assess the behavior of proliferating cells under different conditions, such as drug treatment and genomic editions. Traditionally, growth curves for adherent cells are obtained by seeding the cells in multiple-well plates and counting the total number of cells at different time points. Here, we compare this traditional method to the fluorescence-based method, which is based on the CFSE fluorescence decay over time.
The fluorescence-based method is not dependent on the determination of the total number of cells, but rather is approached by assessing the fluorescence of a sample of single cells from a cell population at different time points after plating. Therefore, this method is not biased due to either cell loss during harvesting or to the presence of cellular debris and cell clumps. Moreover, the fluorescence-based method displays lower variation among different measurements of the same time point, which increases the reliability on the determination of lag, log and stationary phase transitions.
细胞生长曲线是用于分析特定培养条件下体外培养细胞增殖动力学的主要检测方法之一。从细胞生长曲线可以评估增殖细胞在不同条件下的行为，如药物处理和基因组编辑。传统上，贴壁细胞的生长曲线是通过将细胞接种在多孔板中并在不同时间点计数细胞总数来获得的。在此，我们将这种传统方法与基于荧光的方法进行比较，该方法基于CFSE荧光随时间的衰减。
基于荧光的方法不依赖于细胞总数的测定，而是通过评估接种后不同时间点细胞群体中单个细胞样本的荧光来实现。因此，该方法不会因收获过程中的细胞损失或细胞碎片和细胞团块的存在而产生偏差。此外，基于荧光的方法在同一时间点的不同测量之间显示出较低的变异性，这增加了确定延迟期、对数期和稳定期转变的可靠性。
