Ascione Flora, Vasaturo Angela, Caserta Sergio, D'Esposito Vittoria, Formisano Pietro, Guido Stefano
Dipartimento di Ingegneria Chimica dei Materiali e della Produzione Industriale (DICMAPI), Università di Napoli Federico II, P.le Tecchio, 80, 80125 Napoli, Italy.
Dipartimento di Ingegneria Chimica dei Materiali e della Produzione Industriale (DICMAPI), Università di Napoli Federico II, P.le Tecchio, 80, 80125 Napoli, Italy; CEINGE Biotecnologie Avanzate, Via Sergio Pansini, 5, 80131 Naples, Italy; Consorzio Interuniversitario Nazionale per la Scienza e Tecnologia dei Materiali (INSTM), UdR INSTM Napoli Federico II, P.le Tecchio, 80, 80125 Napoli, Italy.
Exp Cell Res. 2016 Sep 10;347(1):123-132. doi: 10.1016/j.yexcr.2016.07.015. Epub 2016 Jul 27.
Cell migration plays a key role in many biological processes, including cancer growth and invasion, embryogenesis, angiogenesis, inflammatory response, and tissue repair. In this work, we compare two well-established experimental approaches for the investigation of cell motility in vitro: the cell random migration (CRM) and the wound healing (WH) assay. In the former, extensive tracking of individual live cells trajectories by time-lapse microscopy and elaborate data processing are used to calculate two intrinsic motility parameters of the cell population under investigation, i.e. the diffusion coefficient and the persistence time. In the WH assay, a scratch is made in a confluent cell monolayer and the closure time of the exposed area is taken as an easy-to-measure, empirical estimate of cell migration. To compare WH and CRM we applied the two assays to investigate the motility of skin fibroblasts isolated from wild type and transgenic mice (TgPED) overexpressing the protein PED/PEA-15, which is highly expressed in patients with type 2 diabetes. Our main result is that the cell motility parameters derived from CRM can be also estimated from a time-resolved analysis of the WH assay, thus showing that the latter is also amenable to a quantitative analysis for the characterization of cell migration. To our knowledge this is the first quantitative comparison of these two widely used techniques.
细胞迁移在许多生物学过程中起着关键作用，包括癌症生长与侵袭、胚胎发育、血管生成、炎症反应和组织修复。在这项工作中，我们比较了两种成熟的体外细胞运动研究实验方法：细胞随机迁移（CRM）和伤口愈合（WH）实验。在前者中，通过延时显微镜对单个活细胞轨迹进行广泛跟踪，并进行精细的数据处理，以计算所研究细胞群体的两个内在运动参数，即扩散系数和持续时间。在WH实验中，在汇合的细胞单层中制造划痕，并将暴露区域的闭合时间作为细胞迁移的易于测量的经验估计值。为了比较WH和CRM，我们应用这两种实验来研究从野生型和过表达蛋白PED/PEA-15的转基因小鼠（TgPED）分离的皮肤成纤维细胞的运动性，该蛋白在2型糖尿病患者中高度表达。我们的主要结果是，从CRM得出的细胞运动参数也可以通过对WH实验的时间分辨分析来估计，从而表明后者也适用于细胞迁移特征的定量分析。据我们所知，这是这两种广泛使用的技术的首次定量比较。
