The matrix-dependent 3D spheroid model of the migration of non-small cell lung cancer: a step towards a rapid automated screening

Abstract

In vitro 3D cell culture systems utilizing multicellular tumor spheroids (MCTS) are widely used in translational oncology, including for studying cell migration and in personalized therapy. However, early stages of cellular migration from MCTS and cross-talk between spheroids are overlooked, which was addressed in the current study. Here, we investigated cell migration from MCTS derived from human non-small cell lung cancer (NSCLC) cell line A549 cultured on different substrates, collagen gel or plastic, at different time points. We found that migration starts at 4–16 h time points after the seeding and its speed is substrate-dependent. We also demonstrated that co-culture of two NSCLC-derived MCTS on collagen gel, but not on plastic, facilitates cell migration compared with single MTCS. This finding should be considered when designing MCTS-based functional assays for personalized therapeutic approach and drug screenings. Overall, our work characterizes the in vitro 3D cell culture model resembling NSCLC cell migration from the clusters of CTCs into surgical wound, and describes microscopy-based tools and approaches for image data analysis with a potential for further automation. These tools and approaches also might be used to predict patterns of CTCs migration based on ex vivo analysis of patient biopsy in a 3D culture system.

A deep learning-based pipeline for analyzing the influences of interfacial mechanochemical microenvironments on spheroid invasion using differential interference contrast microscopic images

“…Exploring the mechanisms behind migratory and invasive behaviors of cancer cells is critical for identifying strategies to counteract metastasis. While the image-based spheroid invasion assay provides many advantages for investigating cancer cell metastasis under in vitro conditions [ 57 , 58 ], the concurrent image processing and analysis during the assay period still present notable challenges. Due to the extensive number of time-lapse brightfield DIC images, the manual processing and analysis of these images is a time-intensive task.…”

Revolutionizing Disease Modeling: The Emergence of Organoids in Cellular Systems

“…They can be generated by culturing cells in non-adherent conditions, such as suspension cultures, hanging drops, or microfluidic devices. Spheroids can be composed of a single cell type or multiple cell types and can be used to study cell–cell interactions, drug screening, and tumor biology [ 112 , 113 , 114 , 115 ].…”

Simple Technique for Microscopic Evaluation of Active Cellular Invasion into 3D Hydrogel Constructs

“…They are not often appropriate for assessing cellular migration into an acellular hydrogel. 27 Scratch assays are a simple and inexpensive motility assessment of cells growing back into an open space in a monolayer. The void in the cell layer can be created by culturing cells around a spacer and subsequently removing the spacer from the culture surface, or by culturing a fully confluent monolayer and then mechanically removing a section of the layer creating a void by scratching.…”

“…However, the use of spheroid cultures generally involves seeding cells within the hydrogel and assessing migration outward from within the hydrogel matrix. They are not often appropriate for assessing cellular migration into an acellular hydrogel …”