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Roswell Park Cancer Institute

6 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Bioluminescence Imaging of NADPH Oxidase Activity in Different Animal Models
Wei Han 1, Hui Li 1, Brahm H. Segal 2,3, Timothy S. Blackwell 1
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.

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Biology

Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes
Joseph D. Tario Jr. 1, Kristen Humphrey 1, Andrew D. Bantly 2, Katharine A. Muirhead 3, Jonni S. Moore 4, Paul K. Wallace 1
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.

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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 Goli 2, Laura S. Weiss 2, Rosie Dau 2, Megha Thomas 2, Shoshanna N. Zucker 2
1Department of Cell Stress Biology, Roswell Park Cancer Institute, 2Department of Pharmaceutical, Social and Administrative Sciences, School of Pharmacy, D'Youville College

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.

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Medicine

Modeling Spontaneous Metastatic Renal Cell Carcinoma (mRCC) in Mice Following Nephrectomy
Amanda Tracz 1, Michalis Mastri 1, Christina R. Lee 2, Roberto Pili 1, John M. L. Ebos 1
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.

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Immunology and Infection

Assessing Anti-fungal Activity of Isolated Alveolar Macrophages by Confocal Microscopy
Melissa J. Grimm 1, Anthony C. D'Auria 1, Brahm H. Segal 2
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.

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Developmental Biology

Generation of Induced Pluripotent Stem Cells from Human Melanoma Tumor-infiltrating Lymphocytes
Hidehito Saito 1,2, Kumiko Iwabuchi 3, Noemi Fusaki 4,5, Fumito Ito 3,6
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.

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