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量化体外划痕试验中实验设计选择的效果。

Quantifying the effect of experimental design choices for in vitro scratch assays.

作者信息

Johnston Stuart T, Ross Joshua V, Binder Benjamin J, Sean McElwain D L, Haridas Parvathi, Simpson Matthew J

机构信息

School of Mathematical Sciences, Queensland University of Technology (QUT), Brisbane, Australia; Institute of Health and Biomedical Innovation, QUT, Brisbane, Australia.

School of Mathematical Sciences, University of Adelaide, Adelaide, Australia.

出版信息

J Theor Biol. 2016 Jul 7;400:19-31. doi: 10.1016/j.jtbi.2016.04.012. Epub 2016 Apr 13.

DOI:10.1016/j.jtbi.2016.04.012
PMID:27086040
Abstract

Scratch assays are often used to investigate potential drug treatments for chronic wounds and cancer. Interpreting these experiments with a mathematical model allows us to estimate the cell diffusivity, D, and the cell proliferation rate, λ. However, the influence of the experimental design on the estimates of D and λ is unclear. Here we apply an approximate Bayesian computation (ABC) parameter inference method, which produces a posterior distribution of D and λ, to new sets of synthetic data, generated from an idealised mathematical model, and experimental data for a non-adhesive mesenchymal population of fibroblast cells. The posterior distribution allows us to quantify the amount of information obtained about D and λ. We investigate two types of scratch assay, as well as varying the number and timing of the experimental observations captured. Our results show that a scrape assay, involving one cell front, provides more precise estimates of D and λ, and is more computationally efficient to interpret than a wound assay, with two opposingly directed cell fronts. We find that recording two observations, after making the initial observation, is sufficient to estimate D and λ, and that the final observation time should correspond to the time taken for the cell front to move across the field of view. These results provide guidance for estimating D and λ, while simultaneously minimising the time and cost associated with performing and interpreting the experiment.

摘要

划痕实验常用于研究慢性伤口和癌症的潜在药物治疗方法。用数学模型解释这些实验可以让我们估计细胞扩散系数(D)和细胞增殖率(\lambda)。然而,实验设计对(D)和(\lambda)估计值的影响尚不清楚。在此,我们将一种近似贝叶斯计算(ABC)参数推断方法应用于从理想化数学模型生成的新的合成数据集以及非粘附间充质成纤维细胞群体的实验数据,该方法可产生(D)和(\lambda)的后验分布。后验分布使我们能够量化获得的关于(D)和(\lambda)的信息量。我们研究了两种类型的划痕实验,以及改变所捕获实验观察的数量和时间。我们的结果表明,涉及一个细胞前沿的刮擦实验能提供更精确的(D)和(\lambda)估计值,并且比具有两个相反方向细胞前沿的伤口实验在计算解释上更高效。我们发现,在进行初始观察后记录两次观察结果就足以估计(D)和(\lambda),并且最终观察时间应与细胞前沿穿过视野所需的时间相对应。这些结果为估计(D)和(\lambda)提供了指导,同时将进行和解释实验相关的时间和成本降至最低。

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