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Lawson Health Research Institute

7 ARTICLES PUBLISHED IN JoVE

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Medicine

Molecular Imaging to Target Transplanted Muscle Progenitor Cells
Kelly Gutpell 1,2, Rebecca McGirr 1, Lisa Hoffman 1,2,3
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.

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Bioengineering

Programming Stem Cells for Therapeutic Angiogenesis Using Biodegradable Polymeric Nanoparticles
Michael Keeney *1, Lorenzo Deveza *1,2, Fan Yang 1,2
1Department of Orthopaedic Surgery, Stanford University , 2Department of Bioengineering, Stanford University

We describe the method of programming stem cells to overexpress therapeutic factors for angiogenesis using biodegradable polymeric nanoparticles. Processes described include polymer synthesis, transfecting adipose-derived stem cells in vitro, and validating the efficacy of programmed stem cells to promote angiogenesis in a murine hindlimb ischemia model.

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Medicine

Adaptation of Semiautomated Circulating Tumor Cell (CTC) Assays for Clinical and Preclinical Research Applications
Lori E. Lowes 1,2, Benjamin D. Hedley 3, Michael Keeney 3,4, Alison L. Allan 1,2,4,5
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.

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Medicine

Quantification of Breast Cancer Cell Invasiveness Using a Three-dimensional (3D) Model
Donna Cvetković *1, Cameron Glenn-Franklin Goertzen *1, Moshmi Bhattacharya 1,2,3
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.

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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. Chu 1,2, Ying Xia 1, Kim Nguyen 1,3, David Goodale 1, Alison Allan 1,2,4,5
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.

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Immunology and Infection

Tailoring In Vivo Cytotoxicity Assays to Study Immunodominance in Tumor-specific CD8+ T Cell Responses
Joshua Choi 1, Courtney E. Meilleur 1, S.M. Mansour Haeryfar 1,2,3,4
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.

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Cancer Research

Isolation and Functional Assessment of Human Breast Cancer Stem Cells from Cell and Tissue Samples
Vasudeva Bhat 1,2, Cory Lefebvre 1,2, David Goodale 1, Mauricio Rodriguez-Torres 1,2, Alison L. Allan 1,2,3,4
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.

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