Time-Series Clustering of Single-Cell Trajectories in Collective Cell Migration

Abstract

Collective invasion drives multicellular cancer cells to spread to surrounding normal tissues. To fully comprehend metastasis, the methodology of analysis of individual cell migration in tissue should be well developed. Extracting and classifying cells with similar migratory characteristics in a colony would facilitate an understanding of complex cell migration patterns. Here, we used electrospun fibers as the extracellular matrix for the in vitro modeling of collective cell migration, clustering of mesenchymal and epithelial cells based on trajectories, and analysis of collective migration patterns based on trajectory similarity. We normalized the trajectories to eliminate the effect of cell location on clustering and used uniform manifold approximation and projection to perform dimensionality reduction on the time-series data before clustering. When the clustering results were superimposed on the trajectories before normalization, the results still exhibited positional similarity, thereby demonstrating that this method can identify cells with similar migration patterns. The same cluster contained both mesenchymal and epithelial cells, and this result was related to cell location and cell division. These data highlight the reliability of this method in identifying consistent migration patterns during collective cell migration. This provides new insights into the epithelial–mesenchymal interactions that affect migration patterns.

Quantitative Analysis of Collective Migration by Single-Cell Tracking Aimed at Understanding Cancer Metastasis

“…Cells before and after induction of EMT by TGF-β exhibited different migratory properties and showed interactions at different ratios. To further analyze this phenomenon of cellular interactions, we propose a method based on a combination of time-series clustering and dimensionality reduction to analyze cells with similar migration patterns [ 26 ]. Whether cell trajectories are related to migration patterns remains unclear.…”