Invasiveness of Cells Leads to Changes in Their Interaction Behavior with the Glycocalyx

Abstract Transendothelial migration is a crucial step during metastasis. Before circulating tumor cells enter the endothelium, they face the glycocalyx. While invasive migration of cancer cells is well studied, few investigations exist regarding their interaction with the glycocalyx. Here, the interaction of three breast cell lines with an endothelial glycocalyx is studied. Benign MCF‐10A, noninvasive malign MCF‐7, and invasive MDA‐MB‐231 cells penetrate the glycocalyx, just adhere to it or approach without even attaching to it. Remarkable fluctuations in these interaction modes are detected by time‐resolved interaction profiles. Adhesion, migration, and invasion characteristics as well as combinations of interaction modes, cell shapes, and cell extensions are studied. The motility and penetration depth into the glycocalyx are analyzed. The invasive cells are the most flexible, penetrating the glycocalyx mostly with a round shape and feet‐like membrane extensions. Noninvasive cancer cells penetrate the glycocalyx the deepest over time and benign cells integrate more likely into the endothelial cell layer underneath the glycocalyx.