Reproducibility of scratch assays is affected by the initial degree of confluence: Experiments, modelling and model selection
Reproducibility of scratch assays is affected by the initial degree of confluence: Experiments, modelling and model selection
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DOI：
10.1016/j.jtbi.2015.10.040
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发表时间：
2016-02-07
影响因子：
2
通讯作者：
Simpson, Matthew J.
中科院分区：
文献类型：
--
作者：
Jin, Wang;Shah, Esha T.;Simpson, Matthew J.
划痕试验难以重现。在这里，我们确定了一个以前被忽视的变异性来源，可以部分解释这一困难。我们分析了一系列划痕试验，其中我们改变了初始融合程度（初始细胞密度）。我们的研究结果表明，再定殖率是非常敏感的初始密度。为了量化细胞迁移和增殖的相对作用，我们将Fisher-Kolmogorov模型的解决方案校准到细胞密度曲线，以提供细胞扩散率D和细胞增殖率λ的估计。这一过程表明，D和A的估计对初始密度非常敏感。这种依赖性表明，费舍尔-柯尔莫哥洛夫模型并不能准确地表示集体细胞扩散过程的细节，因为该模型假设D和λ是常数，应该与初始密度无关。由于较高的初始细胞密度导致增强的扩散，我们还根据数据校准了Porous-Fisher模型的解，因为该模型假设细胞通量是细胞密度的递增函数。与多孔Fisher模型相关的D和λ的估计值对初始密度不太敏感，这表明多孔Fisher模型提供了对实验的更好描述。(C)2015爱思唯尔有限公司版权所有。
Scratch assays are difficult to reproduce. Here we identify a previously overlooked source of variability which could partially explain this difficulty. We analyse a suite of scratch assays in which we vary the initial degree of confluence (initial cell density). Our results indicate that the rate of re-colonisation is very sensitive to the initial density. To quantify the relative roles of cell migration and proliferation, we calibrate the solution of the Fisher-Kolmogorov model to cell density profiles to provide estimates of the cell diffusivity, D, and the cell proliferation rate, lambda. This procedure indicates that the estimates of D and A are very sensitive to the initial density. This dependence suggests that the Fisher-Kolmogorov model does not accurately represent the details of the collective cell spreading process, since this model assumes that D and lambda are constants that ought to be independent of the initial density. Since higher initial cell density leads to enhanced spreading, we also calibrate the solution of the Porous-Fisher model to the data as this model assumes that the cell flux is an increasing function of the cell density. Estimates of D and lambda associated with the Porous-Fisher model are less sensitive to the initial density, suggesting that the Porous-Fisher model provides a better description of the experiments. (C) 2015 Elsevier Ltd. All rights reserved.