Although histological techniques have provided important information on epithelial stem cells and cancer, they draw static images of dynamic processes. To study dynamic processes, we have developed various imaging windows to image intestinal, liver and breast tissue, and visualize the behavior of individual cells at subcellular resolution for several weeks with two-photon intravital microscropy (IVM). Our IVM experiments illustrate that cellular properties and fate of cells are highly dynamic and change over time. For example, we show in healthy and tumorigenic tissues, that cells can acquire and lose stem cell properties, illustrating that stemness is a state as opposed to an intrinsic property of a cell. Moreover, we show that mammary tumor cells that are surrounded by T cells acquire migration properties. An additional aspect that complicates tumor heterogeneity is that cells may exchange active biomolecules through the release and uptake of extracellular vesicles (EVs). Our data shows, in living mice, that malignant tumor cells, through transfer of EVs, enhance the migratory behavior and metastatic capacity of more benign cells. Taken together, these data exemplify that tumor heterogeneity is far more complex than currently anticipated, which has profound consequences for our ideas on the mechanisms of tumor progression and for designing optimal treatment strategies.

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EMC Abstracts - https://emc-proceedings.com/abstract/real-time-subcellular-imaging-of-cancer-cell-behavior-in-living-mice/