Orthotopic Lung Cancer Murine Model by Nonoperative Transbronchial Approach Takahiro Nakajima, MD, PhD, Takashi Anayama, MD, PhD, Yasushi Matsuda, MD,

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Presentation transcript:

Orthotopic Lung Cancer Murine Model by Nonoperative Transbronchial Approach Takahiro Nakajima, MD, PhD, Takashi Anayama, MD, PhD, Yasushi Matsuda, MD, PhD, David M. Hwang, MD, PhD, Patrick Z. McVeigh, Brian C. Wilson, PhD, Gang Zheng, PhD, Shaf Keshavjee, MD, MS, Kazuhiro Yasufuku, MD, PhD The Annals of Thoracic Surgery Volume 97, Issue 5, Pages 1771-1775 (May 2014) DOI: 10.1016/j.athoracsur.2014.01.048 Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

Fig 1 Images taken during tumor inoculation. (A) The mouse was placed in a modified 50-mL centrifuge tube with the head elevated to a 90-degree angle by its 2 front upper teeth with a rubber band. We used Mosquito-pean forceps to lift the lower jaw and expose the glottis. (B) Vocal cords (arrow) were seen through the surgical microscope. (C, D) The 23-gauge, 2.5-cm curved blunt-tip catheter allows us to inoculate tumor suspension to the right or left lower lobe. (E) A roentgenogram shows the catheter in place at the right lower bronchus. The Annals of Thoracic Surgery 2014 97, 1771-1775DOI: (10.1016/j.athoracsur.2014.01.048) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

Fig 2 (A) A series microcomputed tomography (microCT) scans, beginning at tumor inoculation (blue arrow) show in vivo tumor growth (black arrow). (B) The tumor size was measured for each microCT scan. (C) Tumor image by high-frequency (40-MHz) ultrasound imaging. The Annals of Thoracic Surgery 2014 97, 1771-1775DOI: (10.1016/j.athoracsur.2014.01.048) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

Fig 3 The area of the tumor, as visualized on microcomputed tomography, is plotted as a function of time after inoculation in individual mice. At 25 days after inoculation of A549 tumor cells, most of the tumor size reached more than 4 mm diameter in the long axis. The Annals of Thoracic Surgery 2014 97, 1771-1775DOI: (10.1016/j.athoracsur.2014.01.048) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

Fig 4 Examples of images taken under (A) white light and (B) fluorescence of an animal at 28 days after tumor inoculation with the H460-GFP tumor cells at necropsy with the chest cavity open (heart was removed). The tumor is seen growing within the right lower lobe. There was no pleural dissemination or effusion evident in either image. The Annals of Thoracic Surgery 2014 97, 1771-1775DOI: (10.1016/j.athoracsur.2014.01.048) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

Fig 5 Histologic features of the different tumor types: A549 tumor with (A) hematoxylin and eosin (H&E stain; original magnification ×25), (B) elastic trichrome stain (original magnification ×200), and (C) cluster of differentiation (CD)31 stain (original magnification ×200); H460 tumor with (D) H&E stain (original magnification ×100) and (E) elastic trichrome stain (original magnification ×100); and (F) H520 tumor with H&E stain (original magnification ×200). The Annals of Thoracic Surgery 2014 97, 1771-1775DOI: (10.1016/j.athoracsur.2014.01.048) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions

Fig 6 Immunohistochemistry for epidermal growth factor receptor showing different expression patterns between the tumor types, with (A) strong positive expression for A549 (original magnification ×200), (B) weaker expression for H460 (original magnification ×200), and (C) minimal expression for H520 (original magnification ×400). The Annals of Thoracic Surgery 2014 97, 1771-1775DOI: (10.1016/j.athoracsur.2014.01.048) Copyright © 2014 The Society of Thoracic Surgeons Terms and Conditions