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London Health Sciences Centre

5 ARTICLES PUBLISHED IN JoVE

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

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.

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.

Biology

Viral Transgene Expression in Rodent Hearts and the Assessment of Cardiac Arrhythmia Risk

Aizhu Lu^1,2, Jerry Wang^1,2, Ying Xia^1,2, Ruonan Gu¹, Kyoung-Han Kim^1,2, Erin E. Mulvihill^1,3, Darryl R. Davis^1,2, Rob S. Beanlands¹, Wenbin Liang^1,2

¹University of Ottawa Heart Institute, ²Department of Cellular and Molecular Medicine, University of Ottawa, ³Department of Biochemistry, Microbiology and Immunology, University of Ottawa

The present protocol describes methods for transgene expression in rat and mouse hearts by direct intramyocardial injection of the virus under echocardiography guidance. Methods for the assessment of the susceptibility of hearts to ventricular arrhythmias by the programmed electrical stimulation of isolated, Langendorff-perfused hearts are also explained here.