Volume 60, Issue 2, Pages (August 2011)

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Volume 60, Issue 2, Pages 337-343 (August 2011) Real-Time Cancer Cell Tracking by Bioluminescence in a Preclinical Model of Human Bladder Cancer Growth and Metastasis Geertje van der Horst, Joost J. van Asten, Anne Figdor, Christel van den Hoogen, Henry Cheung, Rob F.M. Bevers, Rob C.M. Pelger, Gabri van der Pluijm European Urology Volume 60, Issue 2, Pages 337-343 (August 2011) DOI: 10.1016/j.eururo.2011.05.005 Copyright © 2011 European Association of Urology Terms and Conditions

Fig. 1 UM-UC-3luc2 cells growing in vitro and in a subcutaneous in vivo model. (A) Quantification of biochemically measured luciferase activity and its correlation with cell number in vitro (range: 10–100 000). The R2 coefficient was calculated with SPSS v.17.0. (B) Schematic representation of UM-UC-3luc2 cells subcutaneously injected into a nude mouse. (C) Representative Images 3 h after inoculation with the quantification in relative light units (D). European Urology 2011 60, 337-343DOI: (10.1016/j.eururo.2011.05.005) Copyright © 2011 European Association of Urology Terms and Conditions

Fig. 2 Preclinical model using bioluminescent imaging (BLI) of orthotopically inoculated UM-UC-3luc2 cells. (A) Schematic representation of UM-UC-3luc2 cells orthotopically inoculated with an angiocathether. (B) Total tumor burden; in the insert, the first 9 d are shown. (C) Representative images of a mouse at days 7, 14, and 21 after inoculation. (D) Representative image of a mouse after resection of the bladder tumor. (E) BLI image of a lung with bladder cancer metastases. (F) BLI image of several lymph node metastases. (G) Hematoxylin and eosin (HE)–stained bladder, with magnified insert (J) at day 30. (H) HE-stained lymph node metastasis, with magnified insert (K). (I) HE-stained lung metastasis, with magnified insert (L). (M–O) Representative images of a lung with micrometastases (arrows). European Urology 2011 60, 337-343DOI: (10.1016/j.eururo.2011.05.005) Copyright © 2011 European Association of Urology Terms and Conditions

Fig. 3 Preclinical metastasis model using bioluminescent imaging of UM-UC-3luc2 cells after intracardiac inoculation. (A) Schematic representation of the intracardiac model. (B) Total tumor burden; in the insert, the first 24 d are shown. (C) The number of metastases per mouse. (D) Representative images of a mouse at days 7, 14, 21, and 28 after inoculation. (E and G) Representative images of a Goldner staining showing UM-UC-3luc2 tumor cells in the tibia. (F and H) Magnified inserts, respectively, of Figures 3e and 3 g. B=bone; T=tumor; BM=bone marrow; GP=growth plate. European Urology 2011 60, 337-343DOI: (10.1016/j.eururo.2011.05.005) Copyright © 2011 European Association of Urology Terms and Conditions

Supplementary Fig. 1 Measurement of small UM-UC-3luc2 cell numbers in vitro. (a) Quantification of biochemically measured luciferase activity and its correlation with cell number (curve: 5, 10, 50, 100, 500). The R2 coefficient was calculated with SPSS v.17.0. European Urology 2011 60, 337-343DOI: (10.1016/j.eururo.2011.05.005) Copyright © 2011 European Association of Urology Terms and Conditions

Supplementary Fig. 2 Bioluminescent imaging of UM-UC-3luc2 cells directly after inoculation (orthotopic and intracardiac). (a and b) UM-UC-3luc2 cells were inoculated with an angiocathether via the urethra, and directly afterward, inoculation bioluminescence was measured with the IVIS 100 imaging system: (a) representative images of mice with technical failure such as intraperitoneal inoculation; (b) representative images of mice correctly inoculated with UM-UC-3luc2 cells. (c and d) UM-UC-3luc2 cells were inoculated in the left cardiac ventricle, and 3h after inoculation, bioluminescence was measured with the IVIS 100 imaging system: (c) representative images of mice with pericardial inoculation at day 0 and day 14; (d) representative images of mice correctly inoculated with UM-UC-3luc2 cells at day 0 and day 14. European Urology 2011 60, 337-343DOI: (10.1016/j.eururo.2011.05.005) Copyright © 2011 European Association of Urology Terms and Conditions