Medical University of South Carolina

33 articles published in JoVE

• Neuroscience

  
  The Evoked Potential Operant Conditioning System (EPOCS): A Research Tool and an Emerging Therapy for Chronic Neuromuscular Disorders N. Jeremy Hill^1,2, Disha Gupta^1,2, Amir Eftekhar¹, Jodi A. Brangaccio¹, James J. S. Norton^1,2, Michelle McLeod³, Tim Fake¹, Jonathan R. Wolpaw^1,2,4, Aiko K. Thompson^{3 1}National Center for Adaptive Neurotechnologies, Stratton VA Medical Center, ²Electrical & Computer Engineering Department, State University of New York at Albany, ³College of Health Professions, Medical University of South Carolina, ⁴Department of Biomedical Sciences, State University of New York at Albany The Evoked Potential Operant Conditioning System aids scientific investigation of sensorimotor function and can administer targeted neurobehavioral training that can impact sensorimotor rehabilitation in neuromuscular disorders. This article describes its capabilities and illustrates its application in modifying a simple spinal reflex to achieve lasting improvement in motor function.

• Medicine

  
  A Simplified Model for Heterotopic Heart Valve Transplantation in Rodents Morgan A. Hill¹, Jennie H. Kwon¹, Brielle Gerry¹, Minoo Kavarana¹, Satish N. Nadig*¹, Taufiek K. Rajab*^{1 1}Department of Surgery, Medical University of South Carolina This protocol describes a simple and efficient method for the transplantation of aortic valve leaflets under the renal capsule to allow for the study of alloreactivity of heart valves.

• Cancer Research

  
  Defining Gene Functions in Tumorigenesis by Ex vivo Ablation of Floxed Alleles in Malignant Peripheral Nerve Sheath Tumor Cells Jody Fromm Longo¹, Stephanie N. Brosius^2,3, Steven L. Carroll^{1 1}Department of Pathology and Laboratory Medicine, Medical University of South Carolina, ²Department of Pediatrics and Neurology, Perelman School of Medicine of the University of Pennsylvania, ³Division of Child Neurology, Children's Hospital of Philadelphia Here, we present a protocol for performing gene knockouts that are embryonic lethal in vivo in genetically engineered mouse model-derived tumors and then assessing the effect that the knockout has on tumor growth, proliferation, survival, migration, invasion, and the transcriptome in vitro and in vivo.

• Behavior

  
  Standing Neurophysiological Assessment of Lower Extremity Muscles Post-Stroke John Harvey Kindred^1,2, Christian Finetto³, Jasmine Jamilah Cash³, Mark Goodman Bowden^{1,2,3 1}Department of Research and Development, Ralph H. Johnson Veterans Administration Medical Center, ²College of Health Professions, Department of Rehabilitation Sciences, Division of Physical Therapy, Medical University of South Carolina, ³College of Health Professions, Department of Health Sciences and Research, Medical University of South Carolina This protocol describes the process for performing a neurophysiological assessment of the lower extremity muscles, tibialis anterior and soleus, in a standing position using TMS in people post-stroke. This position provides a greater probability of eliciting a post-stroke TMS response and allows for the use of reduced stimulator power during neurophysiological assessments.

• Medicine

  
  Minimizing Post-Infusion Portal Vein Bleeding during Intrahepatic Islet Transplantation in Mice Wenyu Gou*¹, Wanxing Cui*², Yuki Cui², Hongjun Wang^{1,3 1}Department of Surgery, Medical University of South Carolina, ²Georgetown University, ³Ralph H. Johnson Veterans Affairs Medical Center Here we present refined surgical procedures on successfully performing intraportal islet transplantation, a clinically relevant but technically challenging surgical procedure, in mice.

• Medicine

  
  A Mouse Model for Chronic Pancreatitis via Bile Duct TNBS Infusion Wenyu Gou*¹, Lindsay Swaby*¹, A. Marissa Wolfe², William P. Lancaster^1,2, Katherine A. Morgan¹, Hongjun Wang^{1,2 1}Department of Surgery, Medical University of South Carolina, ²Ralph H. Johnson Veterans Affairs Medical Center Chronic pancreatitis (CP) is a disease characterized by inflammation and fibrosis of the pancreas, often associated with intractable abdominal pain. This article focuses on refining the technique to generate a mouse model of CP via bile duct infusion with 2,4,6 -trinitrobenzene sulfonic acid (TNBS).

• Bioengineering

  
  Encapsulated Cell Technology for the Delivery of Biologics to the Mouse Eye Marwa Belhaj¹, Balasubramaniam Annamalai², Nathaniel Parsons², Andrew Shuler¹, Jay Potts¹, Bärbel Rohrer^{2,3,4 1}Department of Cell Biology, University of South Carolina, ²Department of Ophthalmology, Division of Research, Medical University of South Carolina, ³Department of Neuroscience, Division of Research, Medical University of South Carolina, ⁴Division of Research, Ralph H. Johnson VA Medical Center Presented here is a protocol for the use of alginate as a polymer in microencapsulation of immortalized cells for long-term delivery of biologics to rodent eyes.

• Cancer Research

  
  Rat Mammary Epithelial Cell Transplantation into the Interscapular White Fat Pad Lauren B. Shunkwiler¹, Jill D. Haag², Michael N. Gould², Bart M. G. Smits^{1 1}Department of Pathology and Laboratory Medicine, Medical University of South Carolina, ²Department of Oncology, McArdle Laboratory for Cancer Research, University of Wisconsin-Madison School of Medicine and Public Health This article describes a transplantation method to graft donor rat mammary epithelial cells into the interscapular white fat pad of recipient animals. This method can be used to examine host and/or donor effects on mammary epithelium development and eliminates the need for pre-clearing, thereby extending the usefulness of this technique.

• Biology

  
  Preparation and Utilization of Freshly Isolated Human Detrusor Smooth Muscle Cells for Characterization of 9-Phenanthrol-Sensitive Cation Currents John Malysz¹, Eric S. Rovner², Robert Wake³, Georgi V. Petkov^{1,3,4 1}Department of Pharmaceutical Sciences, College of Pharmacy, University of Tennessee Health Science Center, ²Department of Urology, Medical University of South Carolina, ³Department of Urology, College of Medicine, University of Tennessee Health Science Center, ⁴Department of Pharmacology, College of Medicine, University of Tennessee Health Science Center We describe a method for preparation of the single freshly isolated detrusor smooth muscle cells from human urinary bladder specimens employing a two-step enzymatic procedure. The obtained viable DSM cells can be studied by various single cell techniques including the described amphotericin-B patch-clamp electrophysiology to reveal physiological and pharmacological properties.

• Neuroscience

  
  Cochlear Surface Preparation in the Adult Mouse Qiao-Jun Fang^1,2, Fan Wu¹, Renjie Chai², Su-Hua Sha^{1 1}Department of Pathology and Laboratory Medicine, Medical University of South Carolina, ²MOE Key Laboratory of Developmental Genes and Human Disease, Institute of Life Sciences, Southeast University This article presents a modified cochlear surface preparation method that requires decalcification and use of a cell and tissue adhesive to adhere the pieces of cochlear epithelia to 10 mm round cover slips for immunohistochemistry in adult mouse cochleae.

• Neuroscience

  
  Bilateral Assessment of the Corticospinal Pathways of the Ankle Muscles Using Navigated Transcranial Magnetic Stimulation Charalambos C. Charalambous^1,2, Jing Nong Liang^3,4, Steve A. Kautz^2,5, Mark S. George^5,6, Mark G. Bowden^{2,5,7 1}Department of Neurology, New York University School of Medicine, ²Department of Health Sciences and Research, Medical University of South Carolina, ³Department of Physical Therapy, University of Nevada Las Vegas, ⁴Department of Health Professions, Medical University of South Carolina, ⁵Ralph H. Johnson VA Medical Center, ⁶Department of Psychiatry, Medical University of South Carolina, ⁷Division of Physical Therapy, Medical University of South Carolina The present protocol describes the simultaneous, bilateral assessment of the corticomotor response of the tibialis anterior and soleus during rest and tonic voluntary activation using a single pulse transcranial magnetic stimulation and neuronavigation system.

• Neuroscience

  
  Laboratory Administration of Transcutaneous Auricular Vagus Nerve Stimulation (taVNS): Technique, Targeting, and Considerations Bashar W. Badran*^1,2,3, Alfred B. Yu*², Devin Adair¹, Georgia Mappin³, William H. DeVries³, Dorothea D. Jenkins⁴, Mark S. George^3,5,6, Marom Bikson^{1 1}Department of Biomedical Engineering, City College of New York, ²U.S. Army Research Laboratory, Aberdeen Proving Ground, ³Brain Stimulation Laboratory, Department of Psychiatry, Medical University of South Carolina, ⁴Department of Pediatrics, Medical University of South Carolina, ⁵Department of Neurology, Medical University of South Carolina, ⁶Ralph H. Johnson VA Medical Center A methodological description of the technique, potential targets, and proper administration of transcutaneous auricular vagus nerve stimulation (taVNS) on the human ear is described.

• Developmental Biology

  
  Using Human Induced Pluripotent Stem Cell-derived Hepatocyte-like Cells for Drug Discovery Jui-Tung Liu¹, Mary Paige Lamprecht¹, Stephen A. Duncan^{1 1}Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina The protocol presented here describes a platform for identifying small molecules for the treatment of liver disease. A step-by-step description is presented detailing how to differentiate iPSCs into cells with hepatocyte characteristics in 96-well plates, and to use the cells to screen for small molecules with potential therapeutic activity.

• Bioengineering

  
  Microdissection of Primary Renal Tissue Segments and Incorporation with Novel Scaffold-free Construct Technology Chase A. Arbra*¹, Satish N. Nadig*¹, Sarah Grace Dennis¹, Sanket Pattanaik¹, Heather A. Bainbridge¹, J. Matthew Rhett¹, Stephen A. Fann¹, Carl Atkinson¹, Michael J. Yost^{1 1}Department of Surgery, Medical University of South Carolina Tissue-engineered renal constructs provide a solution for the organ shortage and deleterious effects of dialysis. Here, we describe a protocol to micro dissect murine kidneys for isolation of cortico-medullary segments. These segments are implanted into scaffold-free cellular constructs, forming renal organoids.

• Genetics

  
  Metabolic Labeling and Profiling of Transfer RNAs Using Macroarrays Sophia Emetu*¹, Morgan Troiano*¹, Jacob Goldmintz*¹, Jensen Tomberlin¹, Simon Grelet², Philip H. Howe², Christopher Korey³, Renaud Geslain^{1 1}Laboratory of tRNA Biology, Department of Biology, College of Charleston, ²Department of Biochemistry and Molecular Biology, MUSC, ³Department of Biology, College of Charleston We describe an original transfer RNA analysis platform named SPOt (Streamlined Platform for Observing tRNA). SPOt simultaneously measures cellular levels of all tRNAs in biological samples, in only three steps, and in less than 24 hours.

• Medicine

  
  Clinicopathological Analysis of miRNA Expression in Breast Cancer Tissues by Using miRNA In Situ Hybridization Yogin Patel¹, Ji S. Lee², Hexin Chen^{1 1}Department of Biological Science, Center for Colon Cancer Research, University of South Carolina, ²Department of Pathology, Chonnam National University Medical School Here we present a protocol to detect miRNA expression in breast cancer patient samples using miRNA in situ hybridization.

• Bioengineering

  
  Three-dimensional Biomimetic Technology: Novel Biorubber Creates Defined Micro- and Macro-scale Architectures in Collagen Hydrogels Veronica Rodriguez-Rivera¹, John W. Weidner², Michael J. Yost^{1 1}Department of Surgery - Division of General Surgery, Medical University of South Carolina, ²Department of Chemical Engineering, University of South Carolina An innovative biofabrication technique was developed to engineer three-dimensional constructs that resemble the architectural features, components, and mechanical properties of in vivo tissue. This technique features a newly developed sacrificial material, BSA rubber, which transfers detailed spatial features, reproducing the in vivo architectures of a wide variety of tissues.

• Neuroscience

  
  Quantification of Filamentous Actin (F-actin) Puncta in Rat Cortical Neurons Hailong Li*¹, Marina Aksenova*¹, Sarah J. Bertrand¹, Charles F. Mactutus¹, Rosemarie Booze^{1 1}Laboratory Program in Behavioral, Neuroscience, Department of Psychology, University of South Carolina Filamentous actin (F-actin) plays an important role in spinogenesis, synaptic plasticity, and synaptic stability. Quantification of F-actin puncta is therefore a useful tool to study the integrity of synaptic structures. This protocol describes the procedures of quantifying F-actin puncta labeled with Phalloidin in low-density primary cortical neuronal cultures.

• Medicine

  
  Evaluation of Lung Metastasis in Mouse Mammary Tumor Models by Quantitative Real-time PCR Melissa A. Abt¹, Christina L. Grek², Gautam S. Ghatnekar², Elizabeth S. Yeh^{1 1}Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, ²FirstString Research, Inc. This protocol describes how to use quantitative Real-time PCR (QRT-PCR) to detect tumor cell specific mRNA representing metastasis within the mouse lung tissue.

• Medicine

  
  The Mouse Round-window Approach for Ototoxic Agent Delivery: A Rapid and Reliable Technique for Inducing Cochlear Cell Degeneration Shawn M. Stevens¹, LaShardai N. Brown², Paula C. Ezell³, Hainan Lang^{2 1}Department of Otolaryngology Head and Neck Surgery, Medical University of South Carolina, ²Department of Pathology and Laboratory Sciences, Medical University of South Carolina, ³Intuitive Surgical, Inc Various methods exist for introducing ototoxic agents to the cochleae of animal models. Presented is a surgical protocol for delivery of ototoxic agents to the round window niche. The procedure is reliable, creates targeted intra-cochlear lesions, and avoids mechanical damage to the microarchitecture. Examination of cochlear self-repair/regeneration is possible.

• Bioengineering

  
  Viability of Bioprinted Cellular Constructs Using a Three Dispenser Cartesian Printer Sarah Grace Dennis¹, Thomas Trusk², Dylan Richards³, Jia Jia³, Yu Tan³, Ying Mei³, Stephen Fann¹, Roger Markwald³, Michael Yost^{1 1}Department of Surgery, Medical University of South Carolina, ²Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, ³Department of Bioengineering, Clemson University A Cartesian bioprinter was designed and fabricated to allow multi-material deposition in precise, reproducible geometries, while also allowing control of environmental factors. Utilizing the three-dimensional bioprinter, complex and viable constructs may be printed and easily reproduced.

• Medicine

  
  Induction and Assessment of Ischemia-reperfusion Injury in Langendorff-perfused Rat Hearts Daniel J. Herr¹, Sverre E. Aune¹, Donald R. Menick^{1,2 1}Department of Medicine/Cardiology, Gazes Cardiac Research Institute, Medical University of South Carolina, ²Ralph H. Johnson VA Medical Center, Medical University of South Carolina The isolated rat heart is an enduring model for ischemia reperfusion injury. Here, we describe the process of harvesting the beating heart from a rat via in situ aortic cannulation, Langendorff perfusion of the heart, simulated ischemia-reperfusion injury, and infarct staining to confirm the extent of ischemic insult.

• Developmental Biology

  
  Culture of Embryonic Mouse Cochlear Explants and Gene Transfer by Electroporation Khujista D. Haque¹, Atul K. Pandey¹, Matthew W. Kelley², Chandrakala Puligilla^{1 1}Department of Pathology and Laboratory Medicine, Medical University of South Carolina, College of Medicine, ²Laboratory of Cochlear Development, NIDCD, NIH We present a method that describes isolation and culture of cochlear explants from embryonic mouse inner ear. We also demonstrate a method for gene transfer into cochlear explants via square-wave electroporation. The in vitro explant culture coupled with gene transfer technique enables researchers to study the effects of altering gene expression during development.

• Medicine

  
  A Possible Zebrafish Model of Polycystic Kidney Disease: Knockdown of wnt5a Causes Cysts in Zebrafish Kidneys Liwei Huang¹, An Xiao¹, Andrea Wecker¹, Daniel A. McBride¹, Soo Young Choi², Weibin Zhou³, Joshua H. Lipschutz^{2 1}Department of Medicine, Eastern Virginia Medical School, ²Department of Medicine, Medical University of South Carolina, ³Department of Pediatrics, University of Michigan We describe a method of generating a possible zebrafish model of polycystic kidney disease. We used Tg(wt1b:GFP) fish to visualize kidney structure. Knockdown of wnt5a was by morpholino injection. Pronephric cyst formation after wnt5a knockdown was observed in this GFP transgenic zebrafish.

• Biology

  
  Reconstruction of 3-Dimensional Histology Volume and its Application to Study Mouse Mammary Glands Rushin Shojaii¹, Stephanie Bacopulos^2,3, Wenyi Yang^2,3, Tigran Karavardanyan⁴, Demetri Spyropoulos⁵, Afshin Raouf⁶, Anne Martel^1,4, Arun Seth^{2,3 1}Department of Medical Biophysics, University of Toronto, ²Platform Biological Sciences, Sunnybrook Research Institute, ³Department of Laboratory Medicine and Pathobiology, University of Toronto, ⁴Physical Sciences, Sunnybrook Research Institute, ⁵Department of Pathology and Laboratory Medicine, Medical University of South Carolina, ⁶Manitoba Institute of Cell Biology, University of Manitoba We present an image registration approach for 3-dimensional (3D) histology volume reconstruction, which facilitates the study of the changes of an organ at the level of macrostructures made up of cells . Using this approach, we studied the 3D changes between wild-type and Igfbp7-null mammary glands.

• Medicine

  
  An Orthotopic Bladder Cancer Model for Gene Delivery Studies Laura Kasman¹, Christina Voelkel-Johnson^{1 1}Department of Microbiology & Immunology, Medical University of South Carolina Implantation of cancer cells into the organ of origin can serve as a useful preclinical model to evaluate novel therapies. MB49 bladder carcinoma cells can be grown within the bladder following intravesical instillation. This protocol demonstrates catheterization of the mouse bladder for the purpose of tumor implantation and adenoviral delivery.

• Medicine

  
  A Contusion Model of Severe Spinal Cord Injury in Rats Vibhor Krishna¹, Hampton Andrews¹, Xing Jin², Jin Yu¹, Abhay Varma¹, Xuejun Wen³, Mark Kindy^{1 1}Department of Neuroscience, Division of Neurosurgery, Medical University of South Carolina, ²Bioengineering, Clemson University, ³Clemson-MUSC Bioengineering Joint Program A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

• Medicine

  
  Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model Ningfei An¹, Yubin Kang^{1 1}Division of Hematology-oncology, Department of Medicine, Medical University of South Carolina Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

• Neuroscience

  
  Targeted Labeling of Neurons in a Specific Functional Micro-domain of the Neocortex by Combining Intrinsic Signal and Two-photon Imaging Philip O'Herron¹, Zhiming Shen¹, Zhongyang Lu¹, Adrien E. Schramm¹, Manuel Levy¹, Prakash Kara^{1 1}Department of Neuroscience, Medical University of South Carolina A method is described for labeling neurons with fluorescent dyes in predetermined functional micro-domains of the neocortex. First, intrinsic signal optical imaging is used to obtain a functional map. Then two-photon microscopy is used to label and image neurons within a micro-domain of the map.

• Medicine

  
  Surgical Technique for Spinal Cord Delivery of Therapies: Demonstration of Procedure in Gottingen Minipigs Thais Federici¹, Carl V. Hurtig¹, Kentrell L. Burks¹, Jonathan P. Riley¹, Vibhor Krishna², Brandon A. Miller¹, Eric A. Sribnick¹, Joseph H. Miller³, Natalia Grin¹, Jason J. Lamanna^1,4,5, Nicholas M. Boulis^{1 1}Department of Neurosurgery, Emory University, ²Department of Neuroscience, Medical University of South Carolina, ³Division of Neurosurgery, University of Alabama, Birmingham, ⁴Department of Biomedical Engineering, Georgia Institute of Technology, ⁵Department of Biomedical Engineering, Emory University Short visual description of the surgical technique and device used for the delivery of (gene and cell) therapies into the spinal cord. The technique is demonstrated in the animal but is entirely translatable and currently being used for human application.

• Neuroscience

  
  Dissection of Adult Mouse Utricle and Adenovirus-mediated Supporting-cell Infection Carlene S. Brandon¹, Christina Voelkel-Johnson², Lindsey A. May³, Lisa L. Cunningham^{3 1}Department of Pathology and Laboratory Medicine, Medical University of South Carolina, ²Department of Microbiology & Immunology, Medical University of South Carolina, ³National Institute on Deafness and Other Communication Disorders, National Institutes of Health Mechanosensory hair cells are the receptor cells of the inner ear. The best-characterized in vitro model system of mature mammalian hair cells utilizes organ cultures of utricles from adult mice. We present the dissection of the adult mouse utricle, and we demonstrate adenovirus-mediated infection of supporting cells in cultured utricles.

• Immunology and Infection

  
  'Bioluminescent' Reporter Phage for the Detection of Category A Bacterial Pathogens David A. Schofield¹, Ian J. Molineux², Caroline Westwater^{3 1}BioSciences Division, Guild Associates, Inc., ²Department of Molecular Genetics and Microbiology, University of Texas at Austin, ³Department of Craniofacial Biology, Medical University of South Carolina A simple method for the identification of priority bacterial pathogens is to use genetically engineered reporter phage. These reporter phage, which are specific to their particular host species, are capable of rapidly transducing a bioluminescent signal response to host cells. Herein, we describe the use of reporter phage for the detection of Yersinia pestis.

• Neuroscience

  
  Preparation of Living Isolated Vertebrate Photoreceptor Cells for Fluorescence Imaging Nicholas P. Boyer¹, Chunhe Chen¹, Yiannis Koutalos^{1 1}Storm Eye Institute, Medical University of South Carolina A method is described for the preparation of single living photoreceptor cells from different vertebrate species for fluorescence imaging. The method can be used to image the fluorescence of endogenous fluorophores, such as NADH or vitamin A, or that of exogenously added fluorescent dyes sensitive to Ca²⁺ or other factors.