Schwann Cells

Description

This volume presents an assortment of traditional and emerging experimental procedures relevant to Schwann cell research. The chapters are divided into four parts. Part I contains protocols for in vitro culture, purification, and characterization of primary Schwann cells from diverse species and stages of nerve development. It also contains protocols to create cancer cell lines and engineered Schwann cells from unconventional sources via chemical conversion, induced differentiation or genetic intervention. Parts II and III outline a wide range of methodologies used to study Schwann cells within in vitro and in vivo systems relevant to the analysis of peripheral nerve development, cancer, axon degeneration/regeneration, and myelination. Last but not least, part IV outlines protocols for Schwann cell production, collection, labeling and transplantation in the injured peripheral nerve and spinal cord of experimental animals and human subjects. Authoritative and practical, Schwann Cells: Methods and Protocols aims to aid both experienced and new investigators to make progress in their research endeavors involving Schwann cells. Front Matter ....Pages i-xiii Front Matter ....Pages 1-1 Isolation of Schwann Cell Precursors from Rodents (Rhona Mirsky, Kristjan R. Jessen)....Pages 3-15 Preparation of Neonatal Rat Schwann Cells and Embryonic Dorsal Root Ganglia Neurons for In Vitro Myelination Studies (Patrice Maurel)....Pages 17-37 Isolation and Expansion of Schwann Cells from Transgenic Mouse Models (Jihyun Kim, Haesun A. Kim)....Pages 39-48 Isolation, Culture, and Cryopreservation of Adult Rodent Schwann Cells Derived from Immediately Dissociated Teased Fibers (Natalia D. Andersen, Paula V. Monje)....Pages 49-66 Detailed Protocols for the Isolation, Culture, Enrichment and Immunostaining of Primary Human Schwann Cells (Tamara Weiss, Sabine Taschner-Mandl, Peter F. Ambros, Inge M. Ambros)....Pages 67-86 Magnetic-Activated Cell Sorting for the Fast and Efficient Separation of Human and Rodent Schwann Cells from Mixed Cell Populations (Kristine M. Ravelo, Natalia D. Andersen, Paula V. Monje)....Pages 87-109 Culture and Expansion of Rodent and Porcine Schwann Cells for Preclinical Animal Studies (Adriana E. Brooks, Gagani Athauda, Mary Bartlett Bunge, Aisha Khan)....Pages 111-126 Chemical Conversion of Human Fibroblasts into Functional Schwann Cells (Eva C. Thoma)....Pages 127-136 Derivation of Fate-Committed Schwann Cells from Bone Marrow Stromal Cells of Adult Rats (Y. P. Tsui, Graham K. Shea, Y. S. Chan, Daisy K. Y. Shum)....Pages 137-148 Human Induced Pluripotent Stem Cell-Derived Sensory Neurons for Fate Commitment of Bone Marrow Stromal Cell-Derived Schwann Cells (Sa Cai, Daisy K. Y. Shum, Ying-Shing Chan)....Pages 149-160 Generation and Use of Merlin-Deficient Human Schwann Cells for a High-Throughput Chemical Genomics Screening Assay (Alejandra M. Petrilli, Cristina Fernández-Valle)....Pages 161-173 Front Matter ....Pages 175-175 Lentiviral Transduction of Rat Schwann Cells and Dorsal Root Ganglia Neurons for In Vitro Myelination Studies (Corey Heffernan, Patrice Maurel)....Pages 177-193 Preservation, Sectioning, and Staining of Schwann Cell Cultures for Transmission Electron Microscopy Analysis (Vania W. Almeida, Margaret L. Bates, Mary Bartlett Bunge)....Pages 195-212 Scalable Differentiation and Dedifferentiation Assays Using Neuron-Free Schwann Cell Cultures (Paula V. Monje)....Pages 213-232 The Pseudopod System for Axon-Glia Interactions: Stimulation and Isolation of Schwann Cell Protrusions that Form in Response to Axonal Membranes (Yannick Poitelon, M. Laura Feltri)....Pages 233-253 In Vitro Analysis of the Role of Schwann Cells on Axonal Degeneration and Regeneration Using Sensory Neurons from Dorsal Root Ganglia (Rodrigo López-Leal, Paula Diaz, Felipe A. Court)....Pages 255-267 A Culture Model to Study Neuron-Schwann Cell-Astrocyte Interactions (Susana R. Cerqueira, Yee-Shuan Lee, Mary Bartlett Bunge)....Pages 269-279 Preparation of Matrices of Variable Stiffness for the Study of Mechanotransduction in Schwann Cell Development (Mateusz M. Urbanski, Carmen V. Melendez-Vasquez)....Pages 281-297 Purification of Exosomes from Primary Schwann Cells, RNA Extraction, and Next-Generation Sequencing of Exosomal RNAs (Cristian De Gregorio, Paula Díaz, Rodrigo López-Leal, Patricio Manque, Felipe A. Court)....Pages 299-315 3D Cancer Migration Assay with Schwann Cells (Laura Fangmann, Steffen Teller, Pavel Stupakov, Helmut Friess, Güralp O. Ceyhan, Ihsan Ekin Demir)....Pages 317-325 Front Matter ....Pages 327-327 Teased Fiber Preparation of Myelinated Nerve Fibers from Peripheral Nerves for Vital Dye Staining and Immunofluorescence Analysis (Alejandra Catenaccio, Felipe A. Court)....Pages 329-337 Whole Mount Immunostaining on Mouse Sciatic Nerves to Visualize Events of Peripheral Nerve Regeneration (Xin-Peng Dun, David B. Parkinson)....Pages 339-348 The Use of Low Vacuum Scanning Electron Microscopy (LVSEM) to Analyze Peripheral Nerve Samples (Peter Bond, David B. Parkinson)....Pages 349-357 Analysis of Myelinating Schwann Cells in Human Skin Biopsies (Mario A. Saporta, Renata de Moraes Maciel)....Pages 359-369 Whole Mount In Situ Hybridization and Immunohistochemistry for Zebrafish Larvae (Rebecca L. Cunningham, Kelly R. Monk)....Pages 371-384 Transmission Electron Microscopy for Zebrafish Larvae and Adult Lateral Line Nerve (Rebecca L. Cunningham, Kelly R. Monk)....Pages 385-400 Live Imaging of Schwann Cell Development in Zebrafish (Rebecca L. Cunningham, Kelly R. Monk)....Pages 401-405 Front Matter ....Pages 407-407 Transplantation of Adult Rat Schwann Cells into the Injured Spinal Cord (Ying Dai, Caitlin E. Hill)....Pages 409-438 Schwann Cell Transplantation Methods Using Biomaterials (Christine D. Plant, Giles W. Plant)....Pages 439-453 Viral Transduction of Schwann Cells for Peripheral Nerve Repair (Christine D. Plant, Giles W. Plant)....Pages 455-466 Intraspinal Delivery of Schwann Cells for Spinal Cord Injury (Andrea J. Santamaría, Juan P. Solano, Francisco D. Benavides, James D. Guest)....Pages 467-484 Back Matter ....Pages 485-488