1. Assessment of Proliferation and Cytotoxicity in a Biomimetic Three-Dimensional Model of Lung Cancer 
Luis F. Tapias, MD, Sarah E. Gilpin, PhD, Xi Ren, PhD, Lan Wei, PhD, Bryan C. Fuchs, PhD, Kenneth K. Tanabe, MD, Michael Lanuti, MD, Harald C. Ott, MD 
The Annals of Thoracic Surgery 
Volume 100, Issue 2, Pages (August 2015)
DOI: /j.athoracsur
Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

2. Fig 1
Decellularized whole-lung scaffolds as a model for biomimetic three-dimensional (3D) culture of non-small cell lung cancer (NSCLC) cells. (A) Schematic representation of the workflow to establish biomimetic culture conditions. (B) Rodent decellularized whole-lung scaffold. Note the preserved morphologic features resembling the native organ. (C) Decellularized whole-lung scaffolds inside a bioreactor allowing perfusion of medium to support biomimetic culture. (D) Trans-illuminated SW1573-seeded lung scaffold, showing growth of cancer cells into macroscopic tumor nodules (arrows). (E) Histologic section of SW1573-seeded lung scaffold illustrating the large size of tumor nodules (hematoxylin and eosin stain; scale bar = 1 mm). (H = heart; LL = left lung; PA = pulmonary artery; RL = right lung; T = trachea.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

3. Fig 2
Proliferation of human non-small cell lung cancer (NSCLC) cells lines. (A) Proliferation curves under two-dimensional conditions. (B) Schematic representation of the resazurin reduction perfusion assay. Resazurin-culture medium mixture is perfused through the scaffolds through the PA. Live NSCLC cells within scaffolds metabolize resazurin (blue color) into resorufin (purple-pink color) producing a fluorescent signal. (C) Proliferation curves under biomimetic three-dimensional (3D) culture conditions as determined by the resazurin perfusion assay. (D) Photomicrographs of NSCLC-seeded scaffolds demonstrating absence of toxicity from repeated application of resazurin (i-iii; H&E = hematoxylin and eosin). All cell lines remained highly proliferative under 3D conditions as evidenced by expression of Ki67 (iv-vi; DAPI = 4′,6-diamidino-2-phenylindole stain). (Data presented as means ± SD; scale bars = 100 μm; *p < 0.05.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

4. Fig 3
The resazurin reduction perfusion assay allows for the noninvasive assessment of cytotoxicity. (A) Dose–response curves of the human non-small cell lung cancer cell (NSCLC) lines to cisplatin and erlotinib. (B) Cytotoxic effect of cisplatin (100 μM), and (C) erlotinib (0.2 μM) under biomimetic three-dimensional culture conditions. (D) Gross photographs of NSCLC-seeded scaffolds at the end of the resazurin reduction assay illustrating treatment responses. (IC50 = inhibitory concentration 50; data presented as mean ± SD. #p = 0.08; *p < 0.05.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions

5. Fig 4
Results from the resazurin reduction perfusion assay correlate with markers of treatment effect. (A) Photomicrographs showing dead H358 and PC9 cells after cisplatin and dead PC9 cells after erlotinib, but normal appearance in H358 and SW1573 cells after erlotinib. (B) Expression of caspase 3 showing the same pattern of sensitivity observed with the resazurin reduction assay. (C). Phospho-epidermal growth factor receptor (EGFR) expression showing effective inhibition by erlotinib in PC9 cells and lack of significant expression in SW1573 cells. Scale bars = 100 μm (A) and (B), and 20 μm (C). (DAPI = 4′,6-diamidino-2-phenylindole stain.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2015 The Society of Thoracic Surgeons Terms and Conditions