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. 2019 Mar 28;8(9):911–924. doi: 10.1002/sctm.18-0256

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© 2019 The Authors. stem cells translational medicine published by Wiley Periodicals, Inc. on behalf of AlphaMed Press

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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Internalization of extracellular vesicles/exosomes (EV/Exs) by human microvascular endothelial cells (HMEC) and human umbilical vein endothelial cells (HUVEC) promote migration, growth, and tube formation. (A): Representative Z‐stack projections of immunofluorescence image by Imaris program for carboxyfluorescein N‐succinimidyl ester (CFSE)‐labeled EV/Exs uptake by HUVEC. Orthogonal views are shown with Y–Z and X–Z orientations (left panel). In the right panel, the uptake of CFSE‐labeled EV/Exs by HUVEC and HMEC as assessed by flow cytometry at different time points as indicated. (B): EV/Exs and non‐EV/Exs‐free medium (SN) or medium alone (med) promote HUVEC migration. The %wound closure as monitored on over time as indicated and calculated by ImageJ software. Similar data were obtained with HMEC. (C): Serum‐starved HMEC growth in the absence (med) or presence of EV/Exs or EV/Exs‐free medium (SN) monitored over 48 hours in Incucyte and assessed by integrated software. Similar data were obtained with HUVEC. (D): Organization of HUVEC in tube structures in the absence (med) or presence of EV/Exs or EV/Exs‐free medium (SN) over 24 hours. Representative images of three independent experiments (left panel, ×40, ×100, and ×200 magnification) and histograms presenting number of nodes and junctions as determined by angiogenesis analyzer for ImageJ software (right panel). All results are presented as mean values ± SD from at least two independent experiments. *, p < .05; **, p < .01; ***, p < .001; ****, p < .0001 compared with medium; ^###, p < .001; ^####, p < .0001 compared with SN.