细胞迁移是一种在正常生理过程以及癌症等疾病中发生的基本且复杂的现象。因此，理解细胞迁移在发育生物学和生物医学科学领域非常重要。细胞迁移发生在三个维度（3D）中，并涉及迁移细胞、邻近细胞、细胞外基质和信号分子之间的相互作用。为了理解这一现象，目前大部分可用的技术仍依赖于二维（2D）细胞迁移实验，也被称为划痕实验或创伤愈合实验。这些方法在创建无细胞区域（划痕或伤口）方面存在有限的可重复性。对细胞的机械/热应力是这些方法适用性的另一个问题。为了解决这些问题，我们开发了一种基于3D打印的生物相容性细胞插入物的替代方法，用于在24孔板中定量细胞迁移。通过高通量实验验证了这些插入物，用于跟踪肺癌细胞系（A549细胞系）在标准细胞迁移促进剂和抑制剂存在下的迁移情况。我们还开发了一个相应的图像分析流程，证明了我们的方法在可重复性和简易性等方面胜过了最先进的细胞迁移评估方法。版权所有 © 2023 Joshi, Madhusudanan and Mijakovic.

Cell migration is a fundamental and complex phenomenon that occurs in normal physiology and in diseases like cancer. Hence, understanding cell migration is very important in the fields of developmental biology and biomedical sciences. Cell migration occurs in 3 dimensions (3D) and involves an interplay of migrating cell(s), neighboring cells, extracellular matrix, and signaling molecules. To understand this phenomenon, most of the currently available techniques still rely on 2-dimensional (2D) cell migration assay, also known as the scratch assay or the wound healing assay. These methods suffer from limited reproducibility in creating a cell-free region (a scratch or a wound). Mechanical/heat related stress to cells is another issue which hampers the applicability of these methods. To tackle these problems, we developed an alternative method based on 3D printed biocompatible cell inserts, for quantifying cell migration in 24-well plates. The inserts were successfully validated via a high throughput assay for following migration of lung cancer cell line (A549 cell line) in the presence of standard cell migration promoters and inhibitors. We also developed an accompanying image analysis pipeline which demonstrated that our method outperforms the state-of-the-art methodologies for assessing the cell migration in terms of reproducibility and simplicity.Copyright © 2023 Joshi, Madhusudanan and Mijakovic.