In vivo bio-distribution and homing of endothelial outgrowth cells in a tumour model  Lotte B. Bertelsen, Mette Hagensen, Morten Busk, Rui Zhang, Anne S. Knudsen, Nathalie Nielsen, Lise Falborg, Bjarne K. Møller, Michael R. Horsman, Hans Stødkilde-Jørgensen  Nuclear Medicine and Biology  Volume 41, Issue 10, Pages 848-855 (November 2014) DOI: 10.1016/j.nucmedbio.2014.07.007 Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 1 a EOC viability with and without 111In-tropolone labelling. b Cellular retention of 111In-tropolone at four time points following labelling. Data is presented as mean±1 SE. *Significant difference (P < 0.05) in a one-way ANOVA between the zerohour group and the other groups. Nuclear Medicine and Biology 2014 41, 848-855DOI: (10.1016/j.nucmedbio.2014.07.007) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 2 Cell-bound 111In activity measured in the blood over time. Each time point represents the ratio between radioactivity in the cells and the total radioactivity (cells plus plasma). (Note: the total radioactivity in the blood is less than 4% of the total injected dose at all measured time point). Data is presented as mean±1 SE (n=6). *Significant difference (P < 0.05) in a one way ANOVA between the zerohour group and the other groups. Nuclear Medicine and Biology 2014 41, 848-855DOI: (10.1016/j.nucmedbio.2014.07.007) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 3 Distribution of 111In activity in the blood, lungs, liver, spleen, kidneys and tumour following injection of 111In labelled EOCs. At each time point the activity in the respective organs was calculated as percentage of the injected dose per gram of tissue (% ID/g). Data is presented as mean±1 SE (n=6). *Significant difference (P < 0.05) in a one way ANOVA between the zero hour group and the other groups. Nuclear Medicine and Biology 2014 41, 848-855DOI: (10.1016/j.nucmedbio.2014.07.007) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 4 Comparison of 111In activities in organs following injection of either 111In labelled EOC or injection of free 111In chloride. Data is presented as mean±1 SE (n=5). *Significant difference (P < 0.05) in a Student'st test or Mann–Whitney rank sum test between the labelled EOC group and the 111In chloride group in each organ. Nuclear Medicine and Biology 2014 41, 848-855DOI: (10.1016/j.nucmedbio.2014.07.007) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 5 Representative autoradiography demonstrating the distribution of 111In in the respective organs (a) and in tumours (b) over time. Separate autoradiograms in (a) and (b) cannot be compared directly since intensity is adjusted to optimize visual appearance. Red is most intense and blue is least intense. Nuclear Medicine and Biology 2014 41, 848-855DOI: (10.1016/j.nucmedbio.2014.07.007) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 6 Twenty four hours after injection of CFSE labelled EOCs (green) they were found in lung (a), liver (b), spleen (c) and tumour (d). EOCs were not taken up by macrophages seen by the missing co-localization with a macrophage marker (red). Arrows indicate FITC positive cells; green, CFSE labelled EOCs; red, macrophages; blue, nuclei; a, b, and d: Scale bars 50μm. c, Scale bars 100μm. Nuclear Medicine and Biology 2014 41, 848-855DOI: (10.1016/j.nucmedbio.2014.07.007) Copyright © 2014 Elsevier Inc. Terms and Conditions

Fig. 7 The rim of the tumour was highly vascularized (a and b). EOCs were however not found to be located in the endothelial lining of vessels (c). Arrows indicate FITC positive cells. a, HE stain; b, collagen IV stain (red); c, FITC stain (red). a, Scale bar 200μm. b, Scale bar 100μm. c, Scale bar 50μm. Nuclear Medicine and Biology 2014 41, 848-855DOI: (10.1016/j.nucmedbio.2014.07.007) Copyright © 2014 Elsevier Inc. Terms and Conditions