Lawson Health Research Institute

7 ARTICLES PUBLISHED IN JoVE

Medicine

Molecular Imaging to Target Transplanted Muscle Progenitor Cells

Kelly Gutpell^1,2, Rebecca McGirr¹, 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.

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.

Medicine

Adaptation of Semiautomated Circulating Tumor Cell (CTC) Assays for Clinical and Preclinical Research Applications

Lori E. Lowes^1,2, Benjamin D. Hedley³, 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.

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.

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 Xia¹, Kim Nguyen^1,3, David Goodale¹, 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.

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 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.

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 Goodale¹, 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.