The Research Institute at Nationwide Children's Hospital

7 articles published in JoVE

• Medicine

  
  Successful Orthotopic Liver Transplantation in Mice Utilizing Microcomputed Tomography Angiography Qiang Zeng¹, Doug A. Gouchoe^2,3, Mahboubeh Nabavinia⁴, Yong Gyu Lee², Xi Wang², Terri A. Shaffer⁵, Mitchel R. Stacy^4,5,6, Blake R. Peterson^7,8, Bryan A. Whitson^2,9, Christopher Breuer^1,4, Sylvester M. Black^{1,10 1}Center for Regenerative Medicine, The Research Institute at Nationwide Children’s Hospital, ⁵Division of Vascular Diseases and Surgery, The Ohio State University College of Medicine, ⁶Interdisciplinary Biophysics Graduate Program, The Ohio State University, ⁷Division of Medicinal Chemistry and Pharmacognosy, The Ohio State University College of Pharmacy, ⁸The Ohio State University Comprehensive Cancer Center, ⁹Division of Cardiac Surgery, Department of Surgery, The Ohio State Wexner Medical Center, ¹⁰Division of Transplantation, Department of Surgery, The Ohio State Wexner Medical Center In this protocol, we discuss the implementation of a model of successful orthotopic liver transplantation (OLT) in mice. Additionally, adjuvants to further analyze allograft patency after successful OLT in a mouse are discussed as well, specifically utilizing microcomputed tomography (microCT) scans.

• Neuroscience

  
  In vitro Modeling for Neurological Diseases using Direct Conversion from Fibroblasts to Neuronal Progenitor Cells and Differentiation into Astrocytes Cassandra N. Dennys¹, Julieth A. Sierra-Delgado¹, Shrestha Sinha Ray¹, Annalisa M. Hartlaub¹, Florence S. Roussel¹, Yacidzohara Rodriguez¹, Kathrin Meyer^{1,2 1} We describe a protocol to reprogram human skin-derived fibroblasts into induced Neuronal Progenitor Cells (iNPCs), and their subsequent differentiation into induced Astrocytes (iAs). This method leads to fast and reproducible generation of iNPCs and iAs in large quantities.

• Genetics

  
  Direct Reprogramming of Human Fibroblasts into Myoblasts to Investigate Therapies for Neuromuscular Disorders Camila F. Almeida¹, Emma C. Frair¹, Nianyuan Huang¹, Reid Neinast², Kim L. McBride^2,3,4,6, Robert B. Weiss⁵, Kevin M. Flanigan^1,6, Nicolas Wein^{1,6 1}Center for Cardiovascular Research, The Research Institute at Nationwide Children’s Hospital, ³Division of Genetic and Genomic Medicine, Nationwide Children’s Hospital, ⁵Department of Human Genetics, The University of Utah School of Medicine, ⁶Department of Pediatrics, The Ohio State University This protocol describes the conversion of skin fibroblasts into myoblasts and their differentiation into myotubes. The cell lines are derived from patients with neuromuscular disorders and can be used to investigate pathological mechanisms and to test therapeutic strategies.

• Bioengineering

  
  Seeding and Implantation of a Biosynthetic Tissue-engineered Tracheal Graft in a Mouse Model Matthew G. Wiet*^1,2, Sayali Dharmadhikari*^1,3, Audrey White^1,2, Susan D. Reynolds⁴, Jed Johnson⁵, Christopher K. Breuer^3,6, Tendy Chiang^{1,3 1} Graft stenosis poses a critical obstacle in tissue engineered airway replacement. To investigate cellular mechanisms underlying stenosis, we utilize a murine model of tissue engineered tracheal replacement with seeded bone marrow mononuclear cells (BM-MNC). Here, we detail our protocol, including scaffold manufacturing, BM-MNC isolation, graft seeding, and implantation.

• Medicine

  
  Drug Repurposing Hypothesis Generation Using the "RE:fine Drugs" System Kelly Regan¹, Soheil Moosavinasab², Philip Payne¹, Simon Lin^{2 1}Department of Biomedical Informatics, The Ohio State University, ² Here we describe a protocol using the web-based drug repurposing hypothesis generation tool: "RE:fine Drugs." This protocol can be modified to a user's preferences at the level of the query type (gene, drug or disease) and/or the range of available advanced options.

• Biology

  
  Isolation of Murine Coronary Vascular Smooth Muscle Cells Kathryn E. Husarek^1,2, Xiaojin Zhang², Patricia E. McCallinhart², Pamela A. Lucchesi^2,3, Aaron J. Trask^{2,3 1}School of Biomedical Science, The Ohio State University College of Medicine, ² The purpose of this protocol is to demonstrate the isolation and culture techniques of murine primary vascular smooth muscle cells (VSMCs) from the coronary circulation. Once VSMCs have been isolated, they can be used for many standard culture techniques.

• Medicine

  
  Collection and Extraction of Saliva DNA for Next Generation Sequencing Michael R. Goode¹, Soo Yeon Cheong¹, Ning Li¹, William C. Ray^1,2, Christopher W. Bartlett^{1,3 1} DNA extraction from saliva can provide a readily available source of high molecular weight DNA, with little to no degradation/fragmentation. This protocol provides optimized parameters for saliva collection/storage and DNA extraction to be of sufficient quality and quantity for downstream DNA assays with high quality requirements.