Fig. 3

Multiparametric analysis of EV-hMSC. a Representative FCM dotplots showing the gate protocol for hMSC-EV. The gate of EV was defined by use of microbeads. Standard microbeads with a diameter of 0.9 μm were used to set the upper size limit for the EV and were used to gate the hMSC-EV. hMSC-EV stained with hematopoietic markers (CD34 and CD45) were negative and were positive for hMSC markers (CD90, CD44, CD73) and for EV markers (CD81 and CD63). In gray represents the control (unstained EV) and in black the EV stained with the different antibodies. b Representative dotplot of EV from hMSC-GFP⁺, in these case we first gate the particles that were positive in fluorescence 2 (GFP) and were also smaller than the upper size (0.9 μm). Then we confirmed with these gate that the EV from hMSC-GFP⁺ were positive for mesenchymal markers, negative for hematopoietic markers and positive for CD63 and CD81 (EV markers)