Ohio State University Wexner Medical Center 15 articles published in JoVE Cancer Research Magnetic Fluorescent Bead-Based Dual-Reporter Flow Analysis of PDL1-Vaxx Peptide Vaccine-Induced Antibody Blockade of the PD-1/PD-L1 Interaction Jay Overholser*1, Linlin Guo*1, Pravin T. P. Kaumaya1,2 1Department of Obstetrics & Gynecology, The Ohio State University Wexner Medical Center, 2The James Comprehensive Cancer Center Checkpoint inhibitors are important targets in developing therapies for the battle against cancer. This report introduces a novel PDL1 peptide-based cancer vaccine, PDL1-Vaxx, which induces neutralizing polyclonal antibody production that blocks PD-1/PDL1 complex formation. This work also details the development and testing of a fluorescent bead-based assay for analyzing this activity. Neuroscience Comprehensive Understanding of Inactivity-Induced Gait Alteration in Rodents Junichi Tajino1,2, Tomoki Aoyama1, Hiroshi Kuroki1, Akira Ito1 1Department of Motor Function Analysis, Human Health Sciences, Graduate School of Medicine, Kyoto University, 2Otolaryngology - Head & Neck Surgery, Ohio State Wexner Medical Center The present protocol describes three-dimensional motion tracking/evaluation to depict gait motion alteration of rats after exposure to a simulated disuse environment. Neuroscience Lineage Tracing of Inducible Fluorescently-Labeled Stem Cells in the Adult Mouse Brain Gabriel S. Jensen1,2, Jake W. Willows2, David T. Breault3, Kristy L. Townsend1,2 1Graduate School of Biomedical Science and Engineering, University of Maine, 2Department of Neurosurgery, Ohio State University Wexner Medical Center, 3Boston Children’s Hospital, Harvard Medical School The ability to permanently mark stem cells and their progeny with a fluorophore using an inducible transgenic lineage tracing mouse line allows for spatial and temporal analysis of activation, proliferation, migration, and/or differentiation in vivo. Lineage tracing can reveal novel information about lineage commitment, response to intervention(s), and multipotency. Medicine A Rat Lung Transplantation Model of Warm Ischemia/Reperfusion Injury: Optimizations to Improve Outcomes Yong Gyu Lee1,2, Jung-Lye Kim1,2, Andre F. Palmer3, Brenda F. Reader4, Jianjie Ma1, Sylvester M. Black2,4, Bryan A. Whitson1,2 1Department of Surgery Division of Cardiac Surgery, The Ohio State University Wexner Medical Center, 2The Collaboration for Organ Perfusion, Protection, Engineering and Regeneration (COPPER) Laboratory, The Ohio State University College of Medicine, 3William G. Lowrie Department of Chemical and Biomolecular Engineering, College of Engineering, The Ohio State University, 4Comprehensive Transplant Center, The Ohio State University Wexner Medical Center Here, we present optimizations to a rat lung transplantation model that serve to improve outcomes. We provide a size guide for cuffs based on body weight, a measurement strategy to ascertain the 4th intercostal space, and methods of wound closure and BAL (bronchoalveolar lavage) fluid and tissue collection. Biology Application of Laser Microdissection to Uncover Regional Transcriptomics in Human Kidney Tissue Daria Barwinska1, Michael J. Ferkowicz1, Ying-Hua Cheng1, Seth Winfree1,2, Kenneth W. Dunn1, Katherine J. Kelly1, Timothy A. Sutton1, Brad H. Rovin3, Samir V. Parikh3, Carrie L. Phillips4, Pierre C. Dagher1, Tarek M. El-Achkar1, Michael T. Eadon1, For The Kidney Precision Medicine Project, 1Department of Medicine, Indiana University School of Medicine, 2Department of Cellular & Integrative Physiology, Indiana University School of Medicine, 3Division of Nephrology, Department of Medicine, Ohio State University Wexner Medical Center, 4Division of Pathology, Indiana University School of Medicine We describe a protocol for laser microdissection of sub-segments of the human kidney, including the glomerulus, proximal tubule, thick ascending limb, collecting duct and interstitium. The RNA is then isolated from the obtained compartments and RNA sequencing is carried out to determine changes in the transcriptomic signature within each sub-segment. Cancer Research Pathological Analysis of Lung Metastasis Following Lateral Tail-Vein Injection of Tumor Cells Katie A Thies1, Sarah Steck1, Sue E Knoblaugh2, Steven T Sizemore1 1Department of Radiation Oncology, Arthur G. James Comprehensive Cancer Center and Richard L. Solove Research Institute, The Ohio State University Medical Center, 2Department of Veterinary Biosciences, Comparative Pathology and Digital Imaging Shared Resource, The Ohio State University Intravenous injection of cancer cells is often used in metastasis research, but the metastatic tumor burden can be difficult to analyze. Herein, we demonstrate a tail-vein injection model of metastasis and include a novel approach to analyze the resulting metastatic lung tumor burden. Neuroscience 3D Kinematic Analysis for the Functional Evaluation in the Rat Model of Sciatic Nerve Crush Injury Tianshu Wang1, Akira Ito2, Junichi Tajino1,3, Hiroshi Kuroki2, Tomoki Aoyama1 1Department of Development and Rehabilitation of Motor Function, Human Health Sciences, Graduate School of Medicine, Kyoto University, 2Department of Motor Function Analysis, Human Health Sciences, Graduate School of Medicine, Kyoto University, 3Department of Otolaryngology, The Ohio State University Wexner Medical Center We introduce a kinematic analysis method that uses a three-dimensional motion capture apparatus containing four cameras and data processing software for performing functional evaluations during fundamental research involving rodent models. Medicine Adeno-Associated Virus-Mediated Delivery of CRISPR for Cardiac Gene Editing in Mice Li Xu1, Yandi Gao1, Yeh Siang Lau1, Renzhi Han1 1Department of Surgery, Davis Heart and Lung Research Institute, Biomedical Sciences Graduate Program, Biophysics Graduate Program, Ohio State University Wexner Medical Center Here we provide a detailed protocol to carry out in vivo cardiac gene editing in mice using recombinant Adeno-Associated Virus(rAAV)-mediated delivery of CRISPR. This protocol offers a promising therapeutic strategy to treat dystrophic cardiomyopathy in Duchenne muscular dystrophy and can be used to generate cardiac-specific knockout in postnatal mice. Medicine A Small Animal Model of Ex Vivo Normothermic Liver Perfusion Eliza W. Beal1,2, Curtis Dumond1, Jung-Lye Kim1,2, Clifford Akateh1,2, Emre Eren1, Katelyn Maynard1, Chandan K. Sen3, Jay L. Zweier4, Kenneth Washburn2, Bryan A. Whitson1,3, Sylvester M. Black1,2 1Collaboration for Organ Perfusion, Protection, Engineering and Regeneration (COPPER) Lab, Division of Transplant, Department of Surgery, Comprehensive Transplant Center, Ohio State University Wexner Medical Center, 2Department of Surgery, Division of Transplant, Ohio State University Wexner Medical Center, 3Department of Surgery, Division of CardioThoracic Surgery, Ohio State University Wexner Medical Center, 4Department of Medicine, Ohio State University Wexner Medical Center There is a significant liver donor shortage, and criteria for liver donors have been expanded. Normothermic ex vivo liver perfusion (NEVLP) has been developed to evaluate and modify organ function. This study demonstrates a rat model of NEVLP and tests the ability of pegylated-catalase, to mitigate liver preservation injury. Medicine Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model Eliza W. Beal1, Curtis Dumond1, Jung-Lye Kim1, Khalid Mumtaz1, Don Hayes Jr.1, Ken Washburn1, Bryan A. Whitson1, Sylvester M. Black1 1Collaboration for Organ Perfusion, Protection, Engineering and Regeneration (COPPER) Lab, Division of Transplant, Department of Surgery, Comprehensive Transplant Center, The Ohio State University Wexner Medical Center A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery. Behavior Electrophysiological Motor Unit Number Estimation (MUNE) Measuring Compound Muscle Action Potential (CMAP) in Mouse Hindlimb Muscles W. David Arnold1,2,3, Kajri A. Sheth1, Christopher G. Wier4, John T. Kissel1,3, Arthur H. Burghes1,3,4, Stephen J. Kolb1,3,4 1Department of Neurology, The Ohio State University Wexner Medical Center, 2Department of Physical Medicine and Rehabilitation, The Ohio State University, 3Department of Neuroscience, The Ohio State University Wexner Medical Center, 4Department of Biochemistry and Pharmacology, The Ohio State University Wexner Medical Center We present refined protocols that allow in vivo monitoring of motor unit function in the mouse. Techniques to measure compound muscle action potential (CMAP) and motor unit number estimation (MUNE) in the mouse hind limb muscles innervated by the sciatic nerve are described. Behavior Mindfulness in Motion (MIM): An Onsite Mindfulness Based Intervention (MBI) for Chronically High Stress Work Environments to Increase Resiliency and Work Engagement Maryanna Klatt1, Beth Steinberg2, Anne-Marie Duchemin3 1Department of Family Medicine, The Ohio State University College of Medicine, 2Critical Care Nursing, Wexner Medical Center, 3Department of Psychiatry, Stress, Trauma, and Resilience (STAR) Program, The Ohio State University College of Medicine The Mindfulness in Motion (MIM) protocol offers a pragmatic Mindfulness Based Intervention (MBI) on-site, for persons working in chronically high-stress work environments that significantly increases resiliency and work engagement. The protocol has proven feasible, beneficial, and is easily adaptable to other high-stress workplaces. Medicine Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program Kevin Nelson1,2, Christopher Bobba1,2, Emre Eren3, Tyler Spata4, Malak Tadres2, Don Hayes, Jr.5,6, Sylvester M. Black3,7, Samir Ghadiali*1,2,3, Bryan A. Whitson*2,3,4 1Department of Biomedical Engineering, Ohio State University Wexner Medical Center, 2Davis Heart & Lung Research Institute, Ohio State University Wexner Medical Center, 3The Collaboration for Organ Perfusion, Protection, Engineering and Regeneration (COPPER) Laboratory, Ohio State University Wexner Medical Center, 4Division of Cardiac Surgery, Department of Surgery, Ohio State University Wexner Medical Center, 5Departments of Pediatrics and Internal Medicine, Ohio State University, 6 Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion. Medicine In Vivo Optical Imaging of Brain Tumors and Arthritis Using Fluorescent SapC-DOPS Nanovesicles Zhengtao Chu1,2, Kathleen LaSance3, Victor Blanco1, Chang-Hyuk Kwon5,6, Balveen Kaur5,6, Malinda Frederick4, Sherry Thornton4, Lisa Lemen3, Xiaoyang Qi1,2 1Division of Hematology-Oncology, Department of Internal Medicine, University of Cincinnati College of Medicine, 2Division of Human Genetics, University of Cincinnati College of Medicine, 3Department of Radiology, University of Cincinnati College of Medicine, 4Division of Rheumatology, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, 5Solid Tumor Biology Program, James Comprehensive Cancer Center, The Ohio State University Medical Center, 6Department of Neurological Surgery, James Comprehensive Cancer Center, The Ohio State University Medical Center We describe a multi-angle rotational optical imaging (MAROI) system for in vivo quantitation of a fluorescent marker delivered by saposin C (SapC)-dioleoylphosphatidylserine (DOPS) nanovesicles. Employing mouse models of cancer and arthritis, we demonstrate how the MAROI signal curve analysis can be used for the precise mapping and biological characterization of disease processes. Biology Assessment of Calcium Sparks in Intact Skeletal Muscle Fibers Ki Ho Park1, Noah Weisleder2, Jingsong Zhou3, Kristyn Gumpper1, Xinyu Zhou1, Pu Duann4, Jianjie Ma1, Pei-Hui Lin1 1Department of Surgery, Davis Heart and Lung Research Institute, The Ohio State University Wexner Medical Center, 2Department of Physiology and Cell Biology, Davis Heart and Lung Research Institute, The Ohio State University Wexner Medical Center, 3Department of Molecular Biophysics and Physiology, Rush University Medical Center, 4Department of Internal Medicine, Davis Heart and Lung Research Institute, The Ohio State University Wexner Medical Center Described here is a method to directly measure calcium sparks, the elementary units of Ca2+ release from sarcoplasmic reticulum in intact skeletal muscle fibers. This method utilizes osmotic-stress-mediated triggering of Ca2+ release from ryanodine receptor in isolated muscle fibers. The dynamics and homeostatic capacity of intracellular Ca2+ signaling can be employed to assess muscle function in health and disease.
