Louisiana State University Health Sciences Center 16 articles published in JoVE Biology Isolation, Proliferation and Differentiation of Rhesus Macaque Adipose-Derived Stem Cells Jonquil M. Poret1,2, Patricia E. Molina1,2, Liz Simon1,2 1Department of Physiology, Louisiana State University Health Sciences Center - New Orleans, 2Comprehensive Alcohol-HIV/AIDS Research Center, Louisiana State University Health Sciences Center - New Orleans In this article, we describe isolation of rhesus macaque derived adipose-derived stem cells (ADSCs) using an enzymatic tissue digestion protocol. Next, we describe ADSC proliferation which includes cell detachment, counting and plating. Lastly, ADSC differentiation is described using specific adipogenic inducing agents. Additionally, we describe staining techniques to confirm differentiation. Cancer Research Modeling Breast Cancer in Human Breast Tissue using a Microphysiological System Loren M. Brown1, Katherine L. Hebert2, Rakesh R. Gurrala3, C. Ethan Byrne4, Matthew Burow5, Elizabeth C. Martin4, Frank H. Lau1 1Department of Surgery, Louisiana State University Health Sciences Center, 2Department of Bioinnovation, Tulane University, 3Tulane University School of Medicine, 4Department of Biological and Agricultural Engineering, Louisiana State University, 5Department of Medicine, Tulane University School of Medicine This protocol describes the construction of an in vitro microphysiological system for studying breast cancer using primary human breast tissue with off the shelf materials. Immunology and Infection Isolation, Transfection, and Culture of Primary Human Monocytes Karlie Plaisance-Bonstaff1,2, Celeste Faia2, Dorota Wyczechowska1, Duane Jeansonne1, Cecilia Vittori1,3, Francesca Peruzzi1,2 1Department of Medicine, Louisiana State University Health Sciences Center, 2Department of Microbiology, Immunology, and Parasitology, Louisiana State University Health Sciences Center, 3Department of Biomedical and Clinical Sciences L. Sacco, University of Milan Presented here is an optimized protocol for isolating, culturing, transfecting, and differentiating human primary monocytes from HIV-infected individuals and healthy controls. Biochemistry Preparation of Fungal and Plant Materials for Structural Elucidation Using Dynamic Nuclear Polarization Solid-State NMR Alex Kirui*1, Malitha C. Dickwella Widanage*1, Frederic Mentink-Vigier2, Ping Wang3, Xue Kang1, Tuo Wang1 1Department of Chemistry, Louisiana State University, 2National High Magnetic Field Laboratory, 3Departments of Pediatrics, and Microbiology, Immunology and Parasitology, Louisiana State University Health Sciences Center A protocol for preparing 13C,15N-labeled fungal and plant samples for multidimensional solid-state NMR spectroscopy and dynamic nuclear polarization (DNP) investigations is presented. Behavior A Microphysiologic Platform for Human Fat: Sandwiched White Adipose Tissue Steven D. Scahill1, Maxwell Hunt2, Camille L. Rogers2, Frank H. Lau2 1Department of Pharmacology, Louisiana State University Health Sciences Center, 2Department of Surgery, Louisiana State University Health Sciences Center White adipose tissue (WAT) has critical deficiencies in its current primary culture models, hindering pharmacological development and metabolic studies. Here, we present a protocol to produce an adipose microphysiological system by sandwiching WAT between sheets of stromal cells. This construct provides a stable and adaptable platform for primary WAT culture. Genetics Simultaneous Video-EEG-ECG Monitoring to Identify Neurocardiac Dysfunction in Mouse Models of Epilepsy Vikas Mishra*1, Nicole M. Gautier*1, Edward Glasscock1 1Department of Cellular Biology and Anatomy, Louisiana State University Health Sciences Center Here, we present a protocol to record brain and heart bio signals in mice using simultaneous video, electroencephalography (EEG), and electrocardiography (ECG). We also describe methods to analyze the resulting EEG-ECG recordings for seizures, EEG spectral power, cardiac function, and heart rate variability. Behavior Utilizing the Modified T-Maze to Assess Functional Memory Outcomes After Cardiac Arrest Celeste Y. C. Wu1,2, Francesca M. Lerner4, Alexandre Couto e Silva3, Harlee E. Possoit1,2, Tsung-Han Hsieh1,2, Jake T. Neumann5, Alireza Minagar1, Hung Wen Lin1,2,3, Reggie H. C. Lee1,2 1Department of Neurology, Louisiana State University Health Science Center, 2Center for Brain Health, Louisiana State University Health Science Center, 3Department of Cellular Biology and Anatomy, Louisiana State University Health Science Center, 4Department of Neurology, Cerebral Vascular Disease Research Laboratories, University of Miami Miller School of Medicine, 5Department of Biomedical Sciences, West Virginia School of Osteopathic Medicine This protocol describes the use of a modified T-maze to evaluate functional learning/memory in asphyxia cardiac arrest-induced cerebral ischemia. Biology Detecting Estrogenic Ligands in Personal Care Products using a Yeast Estrogen Screen Optimized for the Undergraduate Teaching Laboratory Thea M. Edwards1,2, Howard E. Morgan2,3, Coralia Balasca4, Naveen K. Chalasani5, Lauren Yam4,6, Alison M. Roark4 1Department of Biology, University of the South, 2School of Biological Sciences, Louisiana Tech University, 3School of Medicine, Louisiana State University Health Sciences Center, 4Department of Biology, Furman University, 5Department of Computer Science, Louisiana Tech University, 6Clemson University This article presents an optimized yeast estrogen screen for quantifying ligands in Personal Care Products (PCPs) that bind estrogen receptors alpha (ERα) and/or beta (ERβ). The method incorporates two colorimetric substrate options, a six-day refrigerated incubation for use in undergraduate courses, and statistical tools for data analysis. Neuroscience Methods and Tips for Intravenous Administration of Adeno-associated Virus to Rats and Evaluation of Central Nervous System Transduction Mychal S. Grames1, Kasey L. Jackson1, Robert D. Dayton1, John A. Stanford2, Ronald L. Klein1 1Department of Pharmacology, Toxicology, and Neuroscience, Louisiana State University Health Sciences Center, 2Department of Molecular & Integrative Physiology, University of Kansas Medical Center Methods for a wide-scale central nervous system gene delivery in the rat are covered. In this example, the purpose is to mimic a disease that affects the entire spinal cord. The widespread transduction can be used to deliver a therapeutic protein to the CNS from a one-time, peripheral administration. Biology The Assembly and Application of 'Shear Rings': A Novel Endothelial Model for Orbital, Unidirectional and Periodic Fluid Flow and Shear Stress Luke A. White1, Emily V. Stevenson1, J. Winny Yun1, Randa Eshaq1, Norman R. Harris1, David K. Mills2, Alireza Minagar3, Pierre-Olivier Couraud4, J. Steven Alexander1 1Molecular and Cellular Physiology, Louisiana State University Health Sciences Center in Shreveport, 2Biological Sciences, Louisiana Tech University, 3Neurology, Louisiana State University Health Sciences Center in Shreveport, 4Institut Cochin, Inserm U1016, Cnrs Umr8104, Université Paris Descartes Different levels and patterns of fluid shear are known to modulate endothelial gene expression, phenotype and susceptibility to disease. We discuss the assembly and use of 'shear rings': a model that produces unidirectional, periodic shear stress patterns. Shear rings are simple to assemble, economical and can produce high cell yields. Medicine Retroductal Submandibular Gland Instillation and Localized Fractionated Irradiation in a Rat Model of Salivary Hypofunction Renjith Parameswaran Nair1, Changyu Zheng2, Gulshan Sunavala-Dossabhoy1 1Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, 2Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, NIH, DHHS Salivary gland hypofunction, a major adverse effect of head and neck radiotherapy diminishes a patient's quality of life. The demonstration of efficacy of new therapies in animal models is a prerequisite before clinical transition. This protocol describes retroductal administration and local irradiation of rat submandibular glands. Medicine Primary Tumor and MEF Cell Isolation to Study Lung Metastasis Shengli Dong*1, Mazvita Maziveyi*1, Suresh K. Alahari1 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. Medicine Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats Rong Jin1, Xiaolei Zhu1, Guohong Li1 1Vascular Biology and Stroke Research Laboratory, Department of Neurosurgery, Louisiana State University Health Science Center, Shreveport In this paper, we demonstrate a standard method for producing an embolic middle cerebral artery occlusion with homologous blood clots (fibrin-rich) in adult rat. This model closely mimics human ischemic stroke and is suitable for preclinical study of thrombolytic therapy for ischemic stroke. Behavior Getting to Compliance in Forced Exercise in Rodents: A Critical Standard to Evaluate Exercise Impact in Aging-related Disorders and Disease Jennifer C. Arnold1, Michael F. Salvatore1 1Department of Pharmacology, Toxicology, & Neuroscience, Louisiana State University Health Sciences Center Exercise may alleviate behavioral impairments associated with neurodegenerative diseases. Studies of the mechanisms underlying exercise-mediated effects have increased in disease models, but the neurobiological background accompanying these diseases, specifically aging, is likely not considered. Here, we illustrate measures to increase the likelihood of a successful exercise paradigm in aged rats. Medicine Intravital Video Microscopy Measurements of Retinal Blood Flow in Mice Norman R. Harris1, Megan N. Watts1, Wendy Leskova1 1Department of Molecular & Cellular Physiology, Louisiana State University Health Sciences Center Intravital microscopy can be used in animals to visualize and measure retinal vascular diameters, bloodstream velocities, and total retinal blood flow. Biology Identifying Protein-protein Interaction in Drosophila Adult Heads by Tandem Affinity Purification (TAP) Xiaolin Tian1, Mingwei Zhu1, Long Li1, Chunlai Wu1 1Neuroscience Center of Excellence, Louisiana State University Health Sciences Center Drosophila is famous for its powerful genetic manipulation, but not for its suitability of in-depth biochemical analysis. Here we present a TAP-based procedure to identify interacting partners of any protein of interest from the fly brain. This procedure can potentially lead to new avenues of research.