JoVE Medicine serves as a connection between basic research and clinical applications. Articles may present medical procedures, case studies or clinical trial methodologies. Included studies are often modeled towards predictive, preventative, personalized, and practical approaches to treatment of disease. The goal of research methodologies in this section is to improve overall patient care and prognosis.
1Department of Baker Institute for Animal Health, Cornell University, 2Department of Molecular Biology and Genetics, Cornell University, 3Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School
Here, we document the use of the soft agar colony formation assay to test the effects of a peptidylarginine deiminase (PADI) enzyme inhibitor, BB-Cl-amidine, on breast cancer tumorigenicity in vitro.
Published May 20, 2015. Keywords: Medicine, Peptidylarginine deiminase enzymes, breast cancer, soft agar assay, cellular transformation, cancer therapies
1Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center
The goal of this protocol is to study breast tumorigenesis. With this technique, mouse mammary tumors are removed and primary cells are prepared from tumors. A lung extraction protocol is included for studying lung metastasis. Furthermore, another protocol for analyzing mouse embryonic fibroblasts from the mouse embryo is included.
Published May 20, 2015. Keywords: Medicine, Tumor, breast, lung, primary, MEF, embryo, fibroblasts, cancer, cell, mouse
1Department of Molecular Biology and Biochemistry, Simon Fraser University
We describe a protocol for aortic interposition grafting in mice. The goal of the protocol is to provide a model with which to study pathological processes and therapeutic strategies relevant to alloimmune reactions in arteries and the resultant arterial changes that contribute to organ transplant failure.
Published May 17, 2015. Keywords: Medicine, Transplantation, Vascular rejection, Transplant arteriosclerosis, Artery, Aorta
1The Perinatal Research Unit, Department of Pediatrics, Institute of Clinical Medicine, Aarhus University Hospital, 2Department of Otorhinolaryngology and Head & Neck Surgery, Institute of Clinical Medicine, Aarhus University Hospital
Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.
Published May 16, 2015. Keywords: Medicine, Piglet, swine, neonatal, hypoxic-ischemic encephalopathy (HIE), asphyxia, hypoxia, amplitude integrated EEG (aEEG), neuroscience, brain injury
1Sargent College, Boston University
Here we demonstrate the most efficient methods for freezing, embedding, cryosectioning, and staining of muscle biopsies to avoid freezing artifacts.
Published May 15, 2015. Keywords: Medicine, skeletal muscle, frozen sections, formaldehyde fixed paraffin embedded sections, cryosectioning, immunohistochemistry, mid belly
1Department of Neurobiology & Anatomy, University of Utah, 2Department of Ophthalmology & Visual Sciences, University of Utah
Microglia activation and microgliosis are key responses to chronic neurodegeneration. Here, we present methods for in vivo, long-term visualization of retinal CX3CR1-GFP+ microglial cells by confocal ophthalmoscopy, and for threshold and morphometric analyses to identify and quantify their activation. We monitor microglial changes during early stages of age-related glaucoma.
Published May 11, 2015. Keywords: Medicine, Neuroscience, microglia, neurodegeneration, glaucoma, retina, optic nerve head, confocal scanning laser ophthalmoscopy, live image analysis, segmentation by thresholding, cell morphometry CX3CR1, DBA/2J
1School of Biomedical Sciences, The University of Queensland
We present here a protocol for the isolation of islets from the mouse model of type 2 diabetes, Leprdb and details of a live-cell assay for measurement of insulin secretion from intact islets that utilizes 2 photon microscopy.
Published May 11, 2015. Keywords: Medicine, Pancreatic islets, Leprdb, db/db, isolation, liberase, collagenase, 2-photon imaging, exocytosis, insulin, beta cell, diabetes
1Department of Medicine, Pulmonary Hypertension Research Group (CRIUCPQ), Laval University, 2Institut National de la Recherche Agronomique, 3Université Diderot Paris, Sorbonne Paris Cité, 4Hôpital Lariboisière, Physiologie clinique Explorations Fonctionnelles, 5INSERM U 965, 6Service de Cardiologie, Centre Hospitalier Universitaire Tours
Pulmonary hypertension is associated with a significant reduction in pulmonary artery pulsatility, contractility and elasticity, contributing to an increase in pulmonary artery pressure and pulmonary resistance. Using a hypoxic piglet model, this study demonstrated that improving pulmonary artery plasticity using a newly developed pulsatile catheter improves hypoxic pulmonary hypertension.
Published May 11, 2015. Keywords: Medicine, Piglets, pulmonary arterial hypertension, right heart catheterization, pulmonary artery pressure, vascular pulsatility, vasodilation, nitric oxide
1Orthopaedic Surgery Department, Hand Surgery Unit, POVISA Hospital, 2Christine M Kleintert Institute for Hand and Microsurgery
Here we describe how to set up a small microsurgical practice station and a simple and inexpensive method for the training of microsurgery with non-living animal models.
Published May 9, 2015. Keywords: Medicine, Micro-surgical, Training, nonliving, chicken, costs, accessible, experimental
1Department of Surgery, Texas A&M University Health Science Center College of Medicine, 2Department of Surgery, Baylor Scott & White Health
Ischemia-Reperfusion (IR) injury is associated with a high rate of morbidity and mortality. The goal of the in vitro model of oxygen-glucose deprivation and reoxygenation (OGD-R) described here is to assess the effects of ischemia reperfusion injury on a variety of cells, particularly in blood-brain barrier (BBB) endothelial cells.
Published May 7, 2015. Keywords: Medicine, Oxygen-glucose deprivation and reoxygenation, ischemia-reperfusion injury, blood-brain barrier, brain endothelial cells, tight junctions, immunofluorescence, f-actin staining