Proceedings of The Physiological Society

Europhysiology 2018 (London, UK) (2018) Proc Physiol Soc 41, PCB067

Poster Communications

Collective and individual dynamics of endothelial cells (HUVEC) in confluent cultures

P. Dieterich1, M. Moskopp1, A. Hohlstamm1, A. Deussen1

1. Institut für Physiologie, Medizinische Fakultät, TU Dresden, Dresden, Germany.

Endothelial cells in confluent monolayers show various signatures of temporal intermittent collective dynamics, e.g. the oriented movement of clusters or the formation of spirals. These effects are the result of a complex interplay of individual active cell dynamics and cell-to-cell interactions. It is the aim of this study to disentangle and quantify these two fundamental aspects. Therefore, we seeded human umbilical vein endothelial cells (HUVEC) on a fibronectin-collagen IV gelatin coated surface, stained the cell nuclei with low concentrations of Hoechst 33342 (1:20000) and observed the dynamics over 24 h with a sampling interval of 10 min. A self-developed image processing software allowed the automated tracking of individual cells resulting in several 10000 cells per time point. An increasing cell number over time showed that cells were viable. In parallel, the mean squared velocity of individual cells as a measure of active motility slowed down over time by about 50 % indicating a non-stationary behavior. In addition, we characterized the dynamics of individual cells from their paths and the corresponding mean squared displacement. We found that endothelial cells showed a normal diffusive behavior with temporal correlations during the first 8 h, but transformed to sub-diffusive dynamics within the following two 8-h-intervals indicating a more persisting cell localization. In parallel, we observed a temporal change of the spatial velocity autocorrelation function of cells indicating changes of interactions between cells. In summary, we have characterized an interesting dynamical behavior of interacting endothelial cells showing anomalous dynamics and complicated time-dependent interaction correlations. In future, this approach will allow to assessing quantitatively the influence of modifications of cell contacts and the cell migration machinery onto the individual and collective dynamics of confluent cells.

Where applicable, experiments conform with Society ethical requirements