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Figure 1 | Journal of Translational Medicine

Figure 1

From: Preferential transfer of mitochondria from endothelial to cancer cells through tunneling nanotubes modulates chemoresistance

Figure 1

Hetero-cellular Tunneling NanoTubes (TnTs) connect stromal to various tumor cells in culture. A, confocal imaging of TnTs in co-culture system. Different tumoral cell lines were co-cultured with eGFP-E4+ECs. Before imaging by confocal microscopy, the co-cultures were stained with Alexa Fluor 594 conjugated-wheat germ agglutinin (WGA AF594) to reveal cell membranes. Hetero-cellular TnTs can be observed between eGFP-E4+ECs in each of the four cell lines studied (MCF7 top left panel, MDA top right panel, SKOV3 bottom left panel, OVCAR3 bottom,right panel). Scale bar: 5 μm. B, confocal imaging of TnTs in co-culture system. MDA, OVCAR3, SKOV3 or MCF7 cells were stained with CellTracker Green, mixed 1/1 with, M-orange-stained Mesenchymal Stem Cells (MSCs) and plated on glass coverslips for subsequent co-cultures. Before imaging, membranes of the co-cultured cells were stained with WGA AF594. Representative images of hetero-cellular TnTs connecting MDA, OVCAR3, SKOV3 or MCF7 cells TnTs to MSCs are presented. Scale bar: 10 μm. C, Phase contrast imaging of ovarian cancer explant. Explants of metastatic ovarian cancer nodules were cultured during ten days in Petri dishes (top left panel). Cells spreading from 3D explants (bottom left panel). TnTs-like structures with high length-to-diameter ratios bridged cells (cell 1 to cell 2 in the right panel). D, confocal imaging of 3D cultures. Angiospheres containing MDA and eGFP-E4+ECs were grown in 3D media for three days and stained with WGA AF594. TnT can be identified between cancer cells in fluorescence (left panel) and in transmission (right panel) in distinct sphere formations. Scale bar: 10 μm. E, confocal imaging of 3D cultures. Angiospheres containing SKOV3 and eGFP-E4+ECs were grown in 3D media for three days and stained with WGA AF594. TnT connecting cancer cells and eGFP-E4+ECs can be observed. Z-X reconstitution shows TnT typical no attachment to the substrate. Scale bar: 10 μm.

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