Indiana University School of Medicine 35 articles published in JoVE Medicine A Reproducible Cartilage Impact Model to Generate Post-Traumatic Osteoarthritis in the Rabbit Julian Dilley*1,2,3, Hessam Noori-Dokht*3,4,6, Abhijit Seetharam1, Margaret Bello1,3, Aaron Nanavaty1,3, Roman M. Natoli2,3, Todd McKinley1,2,3, Zachary Bault5, Diane Wagner2,3,6, Uma Sankar1,3 1Department of Anatomy, Cell Biology and Physiology, Indiana University School of Medicine, 2Department of Orthopaedic Surgery, Indiana University School of Medicine, 3Indiana Center for Musculoskeletal Health, Indiana University School of Medicine, 4School of Mechanical Engineering, Purdue University, 5Large Animal Resource Center, Indiana University School of Medicine, 6Department of Mechanical and Energy Engineering, Indiana University-Purdue University Indianapolis The open medial femoral condyle impact model in rabbits is reliable for studying post-traumatic osteoarthritis (PTOA) and novel therapeutic strategies to mitigate PTOA progression. This protocol generates an isolated cartilage defect of the posterior medial femoral condyle in rabbits using a carriage-based drop tower with an impactor head. Immunology and Infection Swine Model of Biofilm Infection and Invisible Wounds Mohamed El Masry1, Pramod Bhasme1, Shomita S. Mathew-Steiner1, Jessica Smith1, Thomas Smeenge1, Sashwati Roy1, Chandan K. Sen1 1Indiana Center for Regenerative Medicine & Engineering, Indiana University Health Comprehensive Wound Center, Department of Surgery, Indiana University School of Medicine Chronic wounds that are resistant to antibiotics are a major threat to the healthcare system. Biofilm infections are stubborn and hostile and can cause deficient functional wound closure. We report a clinically relevant swine model of biofilm-infected full-thickness chronic wounds. This model is powerful for mechanistic studies as well as for testing interventions. Genetics Adipocyte-Specific ATAC-Seq with Adipose Tissues Using Fluorescence-Activated Nucleus Sorting Kyungchan Kim1, Solaema Taleb1, Jisun So1, Jamie Wann1, Hyun Cheol Roh1 1Department of Biochemistry and Molecular Biology, Indiana University School of Medicine We present a protocol for assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) specifically on adipocytes using nucleus sorting with adipose tissues isolated from transgenic reporter mice with nuclear fluorescence labeling. Behavior The Motivation for Alcohol Reward: Predictors of Progressive-Ratio Intravenous Alcohol Self-Administration in Humans Bethany L. Stangl1, Nia D. Byrd1, Soundarya Soundararajan1, Martin H. Plawecki2, Sean O’Connor2,3, Vijay A. Ramchandani1 1Human Psychopharmacology Laboratory, Division of Intramural Clinical and Biological Research, NIAAA, 2Dept. of Psychiatry, Indiana University School of Medicine, 3R.L. Roudebush VA Medical Center This study aims to show that the Progressive-Ratio Computer-assisted Alcohol-Infusion System (CAIS) paradigm is a reliable and sensitive method that can be used to examine the motivating properties associated with alcohol self-administration in humans. Medicine Using 2-Photon Microscopy to Quantify the Effects of Chronic Unilateral Ureteral Obstruction on Glomerular Processes Mark C, Wagner1, Ruben M. Sandoval1, Silvia B. Campos-Bilderback1, Bruce A. Molitoris1 1Department of Medicine, Division of Nephrology, Indiana University School of Medicine Here, we present a protocol using 2-photon microscopy in Munich Wistar Fromter rats with surface glomeruli to quantifythe effects of prolonged ureteral obstruction on glomerular dynamics and function. Biology Direct Detection of Isolevuglandins in Tissues Using a D11 scFv-Alkaline Phosphatase Fusion Protein and Immunofluorescence Cassandra Warden*1, Alan J. Simmons*2, Lejla Pasic3, Ashley Pitzer4,6, Sean S. Davies4, Justin H. Layer5, Raymond L. Mernaugh3, Annet Kirabo4,6 1Vanderbilt Eye Institute, Vanderbilt University Medical Center, 2Department of Cell and Developmental Biology, Vanderbilt University, 3Department of Biochemistry, Vanderbilt University, 4Division of Clinical Pharmacology, Department of Medicine, Vanderbilt University Medical Center, 5Division of Hematology and Oncology, Indiana University School of Medicine, 6Department of Molecular Physiology and Biophysics, Vanderbilt University This article provides a detailed methodology for the measurement of isolevuglandins in tissues by immunofluorescence using alkaline phosphatase-conjugated ScFv D11 antibody. Hypertension models in both mice and humans are used to explain the step-by-step procedures and fundamental principles associated with isolevuglandin measurement in tissue samples. Biology Artificial Intelligence Approaches to Assessing Primary Cilia Ruchi Bansal1, Staci E. Engle1, Tisianna K. Kamba1, Kathryn M. Brewer1, Wesley R. Lewis2, Nicolas F. Berbari1,3,4 1Department of Biology, Indiana University-Purdue University Indianapolis, 2Nikon Instruments Inc., 3Stark Neurosciences Research Institute, Indiana University, 4Center for Diabetes and Metabolic Diseases, Indiana University School of Medicine The use of artificial intelligence (Ai) to analyze images is emerging as a powerful, less biased, and rapid approach compared with commonly used methods. Here we trained Ai to recognize a cellular organelle, primary cilia, and analyze properties such as length and staining intensity in a rigorous and reproducible manner. Behavior The Participant-Reported Implementation Update and Score (PRIUS): A Novel Method for Capturing Implementation-Related Data Over Time Edward J. Miech1,2,3,4,5, Nicholas A. Rattray1,2,3,6, Dawn M. Bravata1,2,3,5, Jennifer Myers1,3, Teresa M. Damush1,2,3,5 1VA Precision Monitoring (PRIS-M) QUERI, Richard L. Roudebush VA Medical Center, 2William M. Tierney Center for Health Services Research, Regenstrief Institute, 3VA Health Services Research & Development Center for Health Information and Communication (CHIC), Richard L. Roudebush VA Medical Center, 4Department of Emergency Medicine, Indiana University School of Medicine, 5Department of Internal Medicine and Geriatrics, Indiana University School of Medicine, 6Department of Anthropology, Indiana University-Purdue University Indianapolis This protocol describes a novel method for collecting and analyzing data related to ongoing implementation called the Participant-Reported Implementation Update and Score (PRIUS). The PRIUS method allows for the efficient and systematic capture of data over time and from multiple viewpoints in healthcare settings. Medicine A Murine Tail Lymphedema Model Aladdin H. Hassanein*1, Mithun Sinha*1, Colby R. Neumann1, Ganesh Mohan1, Imran Khan1, Chandan K. Sen1 1Department of Surgery, Indiana Center for Regenerative Medicine and Engineering, Indiana University School of Medicine Lymphedema is extremity swelling caused by lymphatic dysfunction. We describe a chronic murine tail model of lymphedema and the novel use of tissue nanotransfection technology (TNT) for genetic cargo delivery to the tail. Medicine Setup and Execution of the Rapid Cycle Deliberate Practice Death Notification Curriculum Patrick G. Hughes1, Kate E. Hughes2, Mary J. Hughes3,4, Lindsay Weaver4, Lauren E. Falvo4, Anna M. Bona4, Dylan Cooper4, Cherri Hobgood4, Rami A. Ahmed4 1Florida Atlantic University Schmidt College of Medicine, 2Department of Emergency Medicine, University of Arizona College of Medicine, 3Department of Osteopathic Medical Specialties, College of Osteopathic Medicine, Michigan State University, 4Indiana University School of Medicine The goal is to demonstrate how to apply the rapid cycle deliberate practice debriefing technique to the GRIEV_ING death notification curriculum. 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. Medicine Leveraging Turbidity and Thromboelastography for Complementary Clot Characterization Ziqian Zeng1,2, Tanmaye Nallan Chakravarthula1,2, Nathan J. Alves1,2 1Emergency Medicine Department, Indiana University School of Medicine, 2Weldon School of Biomedical Engineering, Purdue University Fibrin is responsible for clot formation during hemostasis and thrombosis. Turbidity assays and thromboelastograhy (TEG) can be utilized as synergistic tools that provide complementary assessment of a clot. These two techniques together can give more insight into how clotting conditions affect fibrin clot formation. Medicine Spinal Cord Lateral Hemisection and Asymmetric Behavioral Assessments in Adult Rats Xiao-Jing Lin1,2,3, Shaonan Wen2,6, Ling-Xiao Deng3, Heqiao Dai3, Xiaolong Du3, Chen Chen3,4, Melissa J. Walker3,4, Ting-Bao Zhao1,5, Xiao-Ming Xu3 1Department of Spinal Cord Injury and Repair, Trauma and Orthopedics Institute of Chinese PLA, 960th Hospital, Joint Logistics Support Force of PLA, 2Institute of Military Cognition and Brain Sciences, Academy of Military Medical Sciences, Academy of Military Sciences, 3Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Department of Neurological Surgery and Goodman Campbell Brain and Spine, Indiana University School of Medicine, 4Program in Medical Neuroscience, Paul and Carole Stark Neurosciences Research Institute, Indiana University School of Medicine, 5Department of Orthopedics, the Third Affiliated Hospital, Shandong University, 6Beijing Computing Center, Beijing Academy of Science and Technology Here we describe surgical procedures to produce a reliable spinal cord lateral hemisection (HX) at the 9th thoracic level in adult rats and neurobehavioral assessments designed for detecting asymmetric deficits after such a unilateral injury. Biochemistry Human Peripheral Blood Neutrophil Isolation for Interrogating the Parkinson's Associated LRRK2 Kinase Pathway by Assessing Rab10 Phosphorylation Ying Fan*1, Francesca Tonelli*1, Shalini Padmanabhan2, Marco A.S. Baptista2, Lindsey Riley2, Danielle Smith3, Connie Marras4, Andrew Howden5, Dario R. Alessi1, Esther Sammler1,6 1MRC Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee, 2 Mutations in the leucine rich repeat kinase 2 gene (LRRK2) cause hereditary Parkinson’s disease. We have developed an easy and robust method for assessing LRRK2-controlled phosphorylation of Rab10 in human peripheral blood neutrophils. This may help identify individuals with increased LRRK2 kinase pathway activity. Biology Ex Vivo Method for Assessing the Mouse Reproductive Tract Spontaneous Motility and a MATLAB-based Uterus Motion Tracking Algorithm for Data Analysis Kaley L. Liang*1, Julia O. Bursova*1, Frank Lam1, Xingjuan Chen1, Alexander G. Obukhov1 1The Department of Anatomy, Cell Biology & Physiology, Indiana University School of Medicine Uterine contractions are important for the well-being of females. However, pathologically increased contractility may result in dysmenorrhea, especially in younger females. Here, we describe a simple ex vivo preparation allowing quick assessment of the efficacy of smooth muscle relaxants that may be used for treating dysmenorrhea. Medicine Setup and Execution Of the Blindfolded Code Training Exercise Patrick G. Hughes1, Kate E. Hughes2, Rami A. Ahmed3 1Florida Atlantic University Schmidt College of Medicine, 2Department of Emergency Medicine, University of Arizona College of Medicine, 3Department of Emergency Medicine, Indiana University School of Medicine The blindfolded code training exercise, which blindfolds the team leader in a code resuscitation simulation, is an advanced teaching technique to improve closed-loop communication, organizational skills, and critical thinking. Neuroscience Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice Xiaojing Lin1,2, Tingbao Zhao3, Wenhui Xiong1, Shaonan Wen4, Xiaoming Jin1, Xiao-Ming Xu1 1Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Department of Neurological Surgery and Goodman and Campbell Brain and Spine, Department of Anatomy and Cell Biology, Indiana University School of Medicine, 2Department of Spinal Cord Injury and Repair, Trauma and Orthopedics Institute of Chinese PLA, General Hospital of Jinan Military Region, 3Department of Orthopedics, Shandong Cancer Hospital, Shandong University, 4Department of Neurobiology, Institute of Basic Medical Sciences, Academy Military Medical Sciences We describe an experimental procedure for measuring neuronal activity through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy in vivo. Chemistry Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry Elijah N. McCool*1, Rachele Lubeckyj*1, Xiaojing Shen1, Qiang Kou2, Xiaowen Liu2,3, Liangliang Sun1 1Department of Chemistry, Michigan State University, 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, 3Center for Computational Biology and Bioinformatics, Indiana University School of Medicine A detailed protocol is described for the separation, identification, and characterization of proteoforms in protein samples using capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry (CZE-ESI-MS/MS). The protocol can be used for the high-resolution characterization of proteoforms in simple protein samples and the large-scale identification of proteoforms in complex proteome samples. Biology A Proximal Culture Method to Study Paracrine Signaling Between Cells Subramanyam Dasari1, Taruni Pandhiri1, James Haley1, Dean Lenz2, Anirban K. Mitra1,3,4 1Indiana University School of Medicine, 2Urology, Indiana University Health Southern Indiana Physicians, 3Indiana University Melvin and Bren Simon Cancer Center, 4Department of Medical and Molecular Genetics, Indiana University School of Medicine Paracrine and juxtacrine cellular interactions play an important role in many biological processes, including tumor progression, immune responses, angiogenesis, and development. Here, a proximal culture method is used to study paracrine signaling where the localized concentrations of the secreted factors are maintained while preventing direct cellular contact. Medicine The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing Justin N. Williams1, Yong Li1, Anuradha Valiya Kambrath1, Uma Sankar1 1Department of Anatomy and Cell Biology, Indiana University School of Medicine The murine closed femoral fracture model is a powerful platform to study fracture healing and novel therapeutic strategies to accelerate bone regeneration. The goal of this surgical protocol is to generate unilateral closed femoral fractures in mice using an intramedullary steel rod to stabilize the femur. Immunology and Infection Histological Quantification to Determine Lung Fungal Burden in Experimental Aspergillosis Dylan J. Stolz1, Ethan M. Sands1, Nansalmaa Amarsaikhan1, Angar Tsoggerel1, Steven P. Templeton1 1Department of Microbiology and Immunology, Indiana University School of Medicine - Terre Haute Here we describe a protocol to determine pulmonary fungal burden in mice with invasive aspergillosis by quantification of Gomori's modified methanamine silver staining in histological sections. Use of this method resulted in comparable results with less animals compared to assessment of fungal burden by quantitative PCR of lung fungal DNA. Developmental Biology Kinetic Analysis of Vasculogenesis Quantifies Dynamics of Vasculogenesis and Angiogenesis In Vitro Kaela M. Varberg1,2, Seth Winfree3,4, Kenneth W. Dunn3,4, Laura S. Haneline1,2,5,6,7 1Department of Cellular and Integrative Physiology, Indiana University School of Medicine, 2Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, 3Indiana Center for Biological Microscopy, Indiana University School of Medicine, 4Department of Medicine, Indiana University School of Medicine, 5Department of Pediatrics, Indiana University School of Medicine, 6Department of Microbiology and Immunology, Indiana University School of Medicine, 7Indiana University Simon Cancer Center, Indiana University School of Medicine Here, we present a protocol for time-lapse imaging and analysis of vasculogenesis in vitro using phase contrast microscopy coupled with the open source software, Kinetic Analysis of Vasculogenesis. This protocol can be applied to quantitatively assess the vasculogenic potential of numerous cell types or experimental conditions that model vascular disease. Neuroscience An In Vivo Duo-color Method for Imaging Vascular Dynamics Following Contusive Spinal Cord Injury Chen Chen1,2, Yi Ping Zhang3, Yan Sun1,4, Wenhui Xiong1, Lisa B. E. Shields3, Christopher B. Shields3,5, Xiaoming Jin1, Xiao-Ming Xu1 1Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, and Department of Neurological Surgery, Indiana University School of Medicine, 2Program in Medical Neuroscience, Stark Neurosciences Research Institute, Indiana University School of Medicine, 3Norton Neuroscience Institute, Norton Healthcare, 4Department of Human Anatomy & Histoembryology, School of Basic Medical Sciences, Fudan University, 5Department of Neurological Surgery, University of Louisville School of Medicine We introduce an in vivo imaging method using two different fluorescent dyes to track dynamic spinal vascular changes following a contusive spinal cord injury in adult Sprague-Dawley rats. Medicine A Tissue Displacement-based Contusive Spinal Cord Injury Model in Mice Xiangbing Wu*1, Yi Ping Zhang*2, Wenrui Qu1, Lisa B. E. Shields2, Christopher B. Shields2,3, Xiao-Ming Xu1 1Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Department of Neurological Surgery and Goodman and Campbell Brain and Spine, Indiana University School of Medicine, 2Norton Neuroscience Institute, Norton Healthcare, 3Department of Anatomical Sciences and Neurobiology, University of Louisville We introduce a tissue displacement-based contusive spinal cord injury model that can produce a consistent contusive spinal cord injury in adult mice. Developmental Biology Combining Intravital Fluorescent Microscopy (IVFM) with Genetic Models to Study Engraftment Dynamics of Hematopoietic Cells to Bone Marrow Niches Lin Wang*1, Malgorzata M. Kamocka*2, Amy Zollman3, Nadia Carlesso3 1Wells Center for Pediatric Research, Department of Pediatrics, Indiana University School of Medicine, 2Indiana Center for Biological Microscopy, Department of Medicine, Indiana University School of Medicine, 3Department of Pediatrics, Indiana University School of Medicine Intravital fluorescence microscopy (IVFM) of the calvarium is applied in combination with genetic animal models to study the homing and engraftment of hematopoietic cells into bone marrow (BM) niches. Genetics Measurement of Differentially Methylated INS DNA Species in Human Serum Samples as a Biomarker of Islet β Cell Death Sarah A Tersey1, Jennifer B Nelson1, Marisa M Fisher2, Raghavendra G Mirmira1,3,4 1Department of Pediatrics, IU Center for Diabetes and Metabolic Disease, Indiana University School of Medicine, 2Department of Pediartics, Omaha Children's Hospital and Medical Center, University of Nebraska Medical Center, 3Departments of Biochemistry and Molecular Biology, Medicine, and Cellular and Integrative Physiology, IU Center for Diabetes and Metabolic Disease, Indiana University School of Medicine, 4Indiana Biosciences Research Institute Islet β cell death precedes development of type 1 diabetes, and detecting this process may allow for early therapeutic intervention. Here, we provide a detailed description of how to measure differentially methylated INS DNA species in human serum as a biomarker of β cell death. Cancer Research The Colon-26 Carcinoma Tumor-bearing Mouse as a Model for the Study of Cancer Cachexia Andrea Bonetto1, Joseph E. Rupert1, Rafael Barreto1, Teresa A. Zimmers1 1Department of Surgery, Simon Cancer Center and IUPUI Center for Cachexia Research, Innovation and Therapy, Indiana University School of Medicine Mice bearing the Colon-26 (C26) carcinoma represent a classical model of cancer cachexia. Progressive muscle wasting occurs in association with tumor growth, over-expression of muscle-specific ubiquitin ligases, and reductions in muscle cross-sectional area. Fat loss is also observed. Cachexia is studied in a time-dependent manner with increasing severity of wasting. Biochemistry Directed Protein Packaging within Outer Membrane Vesicles from Escherichia coli: Design, Production and Purification Nathan J. Alves1,2, Kendrick B. Turner1, Scott A. Walper1 1Center for Bio/Molecular Science & Engineering, Naval Research Laboratory, 2Department of Emergency Medicine, Indiana University of School of Medicine A protocol for the production, purification, and use of enzyme packaged outer membrane vesicles (OMV) providing for enhanced enzyme stability for implementation across diverse applications is presented. Biology Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae Xin Zhang*1, Keshava Mysore*2,3, Ellen Flannery3,4, Kristin Michel1, David W. Severson3,4, Kun Yan Zhu5, Molly Duman-Scheel2,3,4 1Division of Biology, Kansas State University, 2Department of Medical and Molecular Genetics, Indiana University School of Medicine, 3Eck Institute for Global Health, University of Notre Dame, 4Department of Biological Sciences, University of Notre Dame, 5Department of Entomology, Kansas State University Here we describe a procedure for inhibiting gene function in disease vector mosquitoes through the use of chitosan/interfering RNA nanoparticles that are ingested by larvae. Medicine Facial Nerve Axotomy in Mice: A Model to Study Motoneuron Response to Injury Deborah N. Olmstead1,2, Nichole A. Mesnard-Hoaglin3, Richard J. Batka1,2, Melissa M. Haulcomb1,2, Whitney M. Miller1,2, Kathryn J. Jones1,2 1Anatomy and Cell Biology, Indiana University School of Medicine, 2Research and Development Services, Richard L. Roudebush VA Medical Center, 3Department of Anatomy and Cell Biology, University of Illinois, Chicago We present a surgical protocol detailing how to perform a cut or crush axotomy on the facial nerve in the mouse. The facial nerve axotomy can be employed to study the physiological response to nerve injury and test therapeutic techniques. Medicine A Novel Vertebral Stabilization Method for Producing Contusive Spinal Cord Injury Melissa J. Walker*1,2, Chandler L. Walker*1,3, Y. Ping Zhang4, Lisa B. E. Shields4, Christopher B. Shields4, Xiao-Ming Xu1,3 1Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Department of Neurological Surgery and Goodman and Campbell Brain and Spine, Indiana University School of Medicine, 2Medical Neuroscience Graduate Program, Indiana University School of Medicine, 3Department of Anatomy and Cell Biology, Indiana University School of Medicine, 4Norton Neuroscience Institute, Norton Healthcare Vertebral stabilization is necessary for minimizing variability, and for producing consistent experimental spinal cord injuries. Using a customized stabilizing apparatus in conjunction with the NYU/MASCIS impactor device, we have demonstrated here the proper equipment and procedure for generating reproducible hemi-contusive cervical (C5) spinal cord injuries in adult rats. Biology Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation Emmanuel Bikorimana1,2, Danica Lapid3, Hyewon Choi3, Richard Dahl1,2,3 1Harper Cancer Research Institute, 2Microbiology and Immunology, Indiana University School of Medicine, 3Department of Biological Sciences, University of Notre Dame Manipulating temporal gene expression in differentiating embryonic stem cells (ESCs) can be achieved using inducible gene systems. However, generation of these cell lines is costly and time consuming. This protocol achieves rapid expression of a transgene in differentiating ES-derived cells and subsequent analysis of downstream hematopoietic differentiation. Medicine Enrichment for Chemoresistant Ovarian Cancer Stem Cells from Human Cell Lines Jennifer M. Cole1, Stancy Joseph1, Christopher G. Sudhahar1, Karen D. Cowden Dahl1 1Department of Biochemistry and Molecular Biology, Indiana University School of Medicine Cancer stem cells (CSCs) are implicated in tumor relapse due to chemoresistance. We have optimized a protocol for selection and expansion of CSCs from ovarian cancer cell lines. By treating cells with the chemotherapeutic cisplatin and culturing in a stem cell promoting media we enrich for non-adherent CSC cultures. Medicine Controlled Cortical Impact Model for Traumatic Brain Injury Jennifer Romine1, Xiang Gao1, Jinhui Chen1 1Spinal Cord and Brain Injury Research Group, Stark Neuroscience Research Institute, Department of Neurological Surgery, Indiana University School of Medicine Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed. Biology qPCR Is a Sensitive and Rapid Method for Detection of Cytomegaloviral DNA in Formalin-fixed, Paraffin-embedded Biopsy Tissue Morgan H. McCoy1, Kristin Post2, Joyashree D. Sen2, Hsim Y. Chang2, Zijin Zhao1, Rong Fan1, Shaoxiong Chen1, Diane Leland1, Liang Cheng1, Jingmei Lin1 1Department of Pathology and Laboratory Medicine, Indiana University School of Medicine, 2Department of Pathology and Laboratory Medicine, Indiana University Health This protocol describes qPCR detection of cytomegalovirus in formalin-fixed, paraffin-embedded biopsy tissue, which is rapid, sensitive, specific, and useful for interpreting equivocal hematoxylin and eosin or immunohistochemical staining patterns.