Sala Federico, Ficorella Carlotta, Martínez Vázquez Rebeca, Eichholz Hannah Marie, Käs Josef A, Osellame Roberto
Department of Physics, Politecnico di Milano, Milan, Italy.
Istituto di Fotonica e Nanotecnologie, Consiglio Nazionale delle Ricerche, Milan, Italy.
Front Bioeng Biotechnol. 2021 Apr 13;9:664094. doi: 10.3389/fbioe.2021.664094. eCollection 2021.
The study of cellular migration dynamics and strategies plays a relevant role in the understanding of both physiological and pathological processes. An important example could be the link between cancer cell motility and tumor evolution into metastatic stage. These strategies can be strongly influenced by the extracellular environment and the consequent mechanical constrains. In this framework, the possibility to study the behavior of single cells when subject to specific topological constraints could be an important tool in the hands of biologists. Two-photon polymerization is a sub-micrometric additive manufacturing technique that allows the fabrication of 3D structures in biocompatible resins, enabling the realization of biochips for cell motility analyses, providing different types of mechanical stimuli. In our work, we present a new strategy for the realization of multilayer microfluidic lab-on-a-chip constructs for the study of cell motility which guarantees complete optical accessibility and the possibility to freely shape the migration area, to tailor it to the requirements of the specific cell type or experiment. The device includes a series of micro-constrictions that induce different types of mechanical stress on the cells during their migration. We show the realization of different possible geometries, in order to prove the versatility of the technique. As a proof of concept, we present the use of one of these devices for the study of the motility of murine neuronal cancer cells under high physical confinement, highlighting their peculiar migration mechanisms.
细胞迁移动力学和策略的研究对于理解生理和病理过程都具有重要意义。一个重要的例子是癌细胞运动性与肿瘤发展到转移阶段之间的联系。这些策略会受到细胞外环境以及随之而来的机械约束的强烈影响。在此框架下，研究单个细胞在特定拓扑约束下的行为对于生物学家而言可能是一项重要工具。双光子聚合是一种亚微米级增材制造技术，可在生物相容性树脂中制造三维结构，从而实现用于细胞运动分析的生物芯片，提供不同类型的机械刺激。在我们的工作中，我们提出了一种实现多层微流控芯片实验室构建体的新策略，用于研究细胞运动，该策略保证了完全的光学可达性以及自由塑造迁移区域的可能性，以使其符合特定细胞类型或实验的要求。该装置包括一系列微缩结构，在细胞迁移过程中会对其施加不同类型的机械应力。我们展示了不同可能几何形状的实现，以证明该技术的通用性。作为概念验证，我们展示了使用其中一种装置研究小鼠神经癌细胞在高度物理限制下的运动性，突出了它们独特的迁移机制。
