Lawson Health Research Institute View Institution's Website 6 articles published in JoVE Cancer Research Isolation and Functional Assessment of Human Breast Cancer Stem Cells from Cell and Tissue Samples Vasudeva Bhat1,2, Cory Lefebvre1,2, David Goodale1, Mauricio Rodriguez-Torres1,2, Alison L. Allan1,2,3,4 1London Regional Cancer Program, 2Department of Anatomy & Cell Biology, Western University, 3Department of Oncology, Western University, 4Lawson Health Research Institute This experimental protocol describes the isolation of BCSCs from breast cancer cell and tissue samples as well as the in vitro and in vivo assays that can be used to assess BCSC phenotype and function. Immunology and Infection Tailoring In Vivo Cytotoxicity Assays to Study Immunodominance in Tumor-specific CD8+ T Cell Responses Joshua Choi1, Courtney E. Meilleur1, S.M. Mansour Haeryfar1,2,3,4 1Department of Microbiology and Immunology, Western University, 2Department of Medicine, Division of Clinical Immunology and Allergy, Western University, 3Department of Surgery, Division of General Surgery, Western University, 4Lawson Health Research Institute We describe here a flow cytometry-based in vivo killing assay that enables examination of immunodominance in cytotoxic T lymphocyte (CTL) responses to a model tumor antigen. We provide examples of how this elegant assay may be employed for mechanistic studies and for drug efficacy testing. Medicine Generation of Organ-conditioned Media and Applications for Studying Organ-specific Influences on Breast Cancer Metastatic Behavior Matthew M. Piaseczny*1,2, Graciella M. Pio*1,2, Jenny E. Chu1,2, Ying Xia1, Kim Nguyen1,3, David Goodale1, Alison Allan1,2,4,5 1London Regional Cancer Program, London Health Sciences Centre, 2Department of Anatomy & Cell Biology, Schulich School of Medicine & Dentistry, Western University, 3Department of Biochemistry, Schulich School of Medicine & Dentistry, Western University, 4Department of Oncology, Schulich School of Medicine & Dentistry, Western University, 5Lawson Health Research Institute This manuscript describes an ex vivo model system comprised of organ-conditioned media derived from the lymph node, bone, lung, and brain of mice. This model system can be used to identify and study organ-derived soluble factors and their effects on the organ tropism and metastatic behavior of cancer cells. Medicine Quantification of Breast Cancer Cell Invasiveness Using a Three-dimensional (3D) Model Donna Cvetković*1, Cameron Glenn-Franklin Goertzen*1, Moshmi Bhattacharya1,2,3 1Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, University of Western Ontario, 2Department of Oncology, Schulich School of Medicine and Dentistry, University of Western Ontario, 3Lawson Health Research Institute This article provides detailed methodologies for the use of three-dimensional (3D) assays to quantify breast cancer cell invasion. Specifically, we discuss the procedures required to set up such assays, quantification, and data analysis, as well as methods to examine the loss of membrane integrity that occurs when cells invade. Medicine Adaptation of Semiautomated Circulating Tumor Cell (CTC) Assays for Clinical and Preclinical Research Applications Lori E. Lowes1,2, Benjamin D. Hedley3, Michael Keeney3,4, Alison L. Allan1,2,4,5 1London Regional Cancer Program, London Health Sciences Centre, 2Department of Anatomy & Cell Biology, Schulich School of Medicine and Dentistry, Western University, 3Special Hematology/Flow Cytometry, London Health Sciences Centre, 4Lawson Health Research Institute, 5Department of Oncology, Western University Circulating tumor cells (CTCs) are prognostic in several metastatic cancers. This manuscript describes the gold standard CellSearch system (CSS) CTC enumeration platform and highlights common misclassification errors. In addition, two adapted protocols are described for user-defined marker characterization of CTCs and CTC enumeration in preclinical mouse models of metastasis using this technology. Medicine Molecular Imaging to Target Transplanted Muscle Progenitor Cells Kelly Gutpell1,2, Rebecca McGirr1, Lisa Hoffman1,2,3 1Imaging Program, Lawson Health Research Institute, 2Department of Anatomy and Cell Biology, Western University, 3Department of Medical Biophysics, Western University A non-invasive means to evaluate the success of myoblast transplantation is described. The method takes advantage of a unified fusion reporter gene composed of genes whose expression can be imaged with different imaging modalities. Here, we make use of a fluc reporter gene sequence to target cells via bioluminescence imaging.