Roswell Park Cancer Institute View Institution's Website 7 articles published in JoVE Developmental Biology Generation of Induced Pluripotent Stem Cells from Human Melanoma Tumor-infiltrating Lymphocytes Hidehito Saito1,2, Kumiko Iwabuchi3, Noemi Fusaki4,5, Fumito Ito3,6 1Department of Surgery, University of Michigan, 2Department of Biochemistry II, Kanazawa Medical University, 3Center for Immunotherapy, Roswell Park Cancer Institute, 4DNAVEC Corporation, 5Department of Ophthalmology, Keio University School of Medicine, 6Department of Surgical Oncology, Roswell Park Cancer Institute The goal of this protocol is to show the protocol for reprogramming melanoma tumor-infiltrating lymphocytes into induced pluripotent stem cells. Medicine A Modified In vitro Invasion Assay to Determine the Potential Role of Hormones, Cytokines and/or Growth Factors in Mediating Cancer Cell Invasion Archis Bagati*1, Zethan Koch*2, Diane Bofinger*2, Haneesha Goli2, Laura S. Weiss2, Rosie Dau2, Megha Thomas2, Shoshanna N. Zucker2 1Department of Cell Stress Biology, Roswell Park Cancer Institute, 2 This video article describes a modified in vitro method to examine the role of hormones, cytokines and/or growth factors in driving cancer cell invasion. Medicine Substernal Thyroid Biopsy Using Endobronchial Ultrasound-guided Transbronchial Needle Aspiration Abhishek Kumar1, Arjun Mohan1, Samjot S. Dhillon2, Kassem Harris2 1Division of Pulmonary and Critical Care Medicine, State University of New York, Buffalo, 2Interventional Pulmonology Section, Division of Medicine, Roswell Park Cancer Institute, State University of New York, Buffalo Substernal thyroid lesions are common, and need to be differentiated from malignancy. Obtaining percutaneous fine needle biopsy is not possible due to its retrosternal location. This article proposes a protocol for biopsy of substernal thyroid lesions using Endobronchial Ultrasound-guided Transbronchial Needle Aspiration (EBUS-TBNA). Immunology and Infection Assessing Anti-fungal Activity of Isolated Alveolar Macrophages by Confocal Microscopy Melissa J. Grimm1, Anthony C. D'Auria1, Brahm H. Segal2 1Department of Medicine, Roswell Park Cancer Institute, 2Department of Medicine, School of Medicine and Biomedical Sciences, University of Buffalo A method to evaluate the ability of isolated mouse alveolar macrophages to control the growth of phagocytosed Aspergillus spores by confocal microscopy. Medicine Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy Amanda Tracz1, Michalis Mastri1, Christina R. Lee2, Roberto Pili1, John M. L. Ebos1 1Genitourinary Section, Department of Medicine, Roswell Park Cancer Institute, 2Biological Sciences Platform, Sunnybrook Research Institute Models of spontaneous metastatic renal cell carcinoma (RCC) disease progression can be used for evaluating treatments in a clinically relevant setting. This protocol demonstrates different procedures for orthotopic kidney tumor cell implantation, proper nephrectomy, and finally outlines a necropsy guide for visual and bioluminescent scoring of metastatic burden and localization. Biology Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes Joseph D. Tario Jr.1, Kristen Humphrey1, Andrew D. Bantly2, Katharine A. Muirhead3, Jonni S. Moore4, Paul K. Wallace1 1Department of Flow and Image Cytometry, Roswell Park Cancer Institute, 2Flow Cytometry & Cell Sorting Resource Laboratory, University of Pennsylvania, 3SciGro, Inc., 4Department of Pathology and Laboratory Medicine, University of Pennsylvania Successful use of cell tracking dyes to monitor immune cell function and proliferation involves several critical steps. We describe methods for: 1) obtaining bright, uniform, reproducible label-ing with membrane dyes; 2) selecting fluorochromes and data acquisition conditions; and 3) choosing a model to quantify cell proliferation based on dye dilution. Immunology and Infection Bioluminescence Imaging of NADPH Oxidase Activity in Different Animal Models Wei Han1, Hui Li1, Brahm H. Segal2,3, Timothy S. Blackwell1 1Department of Medicine, Vanderbilt University School of Medicine, 2Departments of Medicine and Immunology, Roswell Park Cancer Institute, 3Department of Medicine, University at Buffalo School of Medicine NADPH oxidase is the major source of reactive oxygen species (ROS) in phagocytes. Because of the ephemeral nature of ROS, it is difficult to measure and monitor ROS levels in living animals. A minimally invasive method for serial quantification of ROS in living mice is described.