Schwann Cells, 1st ed. 2018 Methods and Protocols Methods in Molecular Biology Series, Vol. 1739

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.

PART I :  Protocols for preparing primary cultures, cell lines and engineered Schwann cells

1. Isolation of Schwann Cell Precursors from Rodents
Rhona Mirsky and Kristjan R. Jessen

2. Preparation of Neonatal Rat Schwann cells and Embryonic Dorsal Root Ganglia Neurons for In Vitro Myelination Studies

Patrice Maurel

3. Isolation and Expansion of Schwann Cells from Transgenic Mouse Models
Jihyun Kim and Haesun Kim

4. Isolation, Culture, and Cryopreservation of Adult Rodent Schwann Cells Derived from Immediately Dissociated Teased Fibers

Natalia D. Andersen and Paula V. Monje

5. Detailed Protocols for the Isolation, Culture, Enrichment, and Immunostaining of Primary Human Schwann cells

6. 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, and Paula V. Monje

7. Culture and Expansion of Rodent and Porcine Schwann Cell for Pre-clinical Animal Studies

Adriana E. Brooks, Gagani Athauda, Mary Bartlett Bunge, and Aisha Khan

8. Chemical Conversion of Human Fibroblasts into functional Schwann Cells

Eva C. Thoma

9. Derivation of Fate-committed Schwann Cells from Bone Marrow Stromal Cells of Adult Rats

Y.P. Tsui, Graham K. Shea, Y.S. Chan, and Daisy K. Shum

10. Human Induced Pluripotent Stem Cell (hiPS cell)-derived Sensory Neurons for Fate Commitment of Bone Marrow Stromal Cell-derived Schwann Cells

Sa Cai, Daisy KY Shum, and Ying-Shing Chan 

11. Generation and Use of Merlin-Deficient Human Schwann Cells for a High-Throughput Chemical Genomics Screening Assay
Alejandra M. Petrilli and Cristina Fernández-Valle

PART II: Protocols for studying Schwann cells behavior and myelination in vitro

12. Lentiviral Transduction of Rat Schwann Cells and Dorsal Root Ganglia Neurons for In Vitro Myelination Studies

Corey Heffernan and Patrice Maurel

Vania W. Almeida, Margaret L. Bates, and Mary Bartlett Bunge

14. Scalable Differentiation and Dedifferentiation Assays using Neuron-free Schwann Cell Cultures

Paula V. Monje

15. The Pseudopod System for Axon-glia Interactions: Stimulation and Isolation of Schwann cell Protrusions that form in Response to Axonal Membranes

Yannick Poitelon and M. Laura Feltri

16. 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, and Felipe A. Court

Susana R. Cerqueira, Yee-Shuan Lee, and Mary Bartlett Bunge

18. Preparation of Matrices of Variable Stiffness for the Study of Mechanotransduction in Schwann Cell Development
Mateusz M. Urbanski and Carmen V. Melendez-Vasquez

19. 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, and Felipe A. Court

20. 3D Cancer Migration Assay with Schwann Cells

Laura Fangmann, Steffen Teller, Pavel Stupakov, Helmut Friess, Güralp O. Ceyhan, and Ihsan Ekin Demir

PART III:  Protocols for studying Schwann cell development and myelination in vivo

21. Teased Fiber Preparation of Myelinated Nerve Fibers from Peripheral Nerves for Vital Dye Staining and Immunofluorescence Analysis

Alejandra Catenaccio and Felipe A. Court

22. Whole Mount Immunostaining on Mouse Sciatic Nerves to Visualize Events of Peripheral Nerve Regeneration
Xin-peng Dun and David B. Parkinson

23. The use of Low Vacuum Scanning Electron Microscopy (LVSEM) to Analyze Peripheral Nerve Myelin
Peter Bond and David Parkinson

24. Analysis of Myelinating Schwann Cells in Human Skin Biopsies

Mario A. Saporta and Renata de Moraes Maciel

25. Whole Mount In Situ Hybridization and Immunohistochemistry for Myelin in Zebra Fish Larvae

Rebecca L. Cunningham and Kelly R. Monk

26. Transmission Electron Microscopy for Zebrafish Larvae and Adult Lateral Line Nerve to Visualize Myelin
Rebecca L. Cunningham and Kelly R. Monk

27. Live Imaging of Schwann Cell Development in Zebra Fish
Rebecca L. Cunningham and Kelly R. Monk

PART IV: Protocols for Schwann cell transplantation

28. Transplantation of Adult Rat Schwann Cells into the Injured Spinal Cord

Ying Dai and Caitlin E. Hill

29. Schwann Cell Transplantation Methods Using Biomaterials

Christine D. Plant and Giles W. Plant

Christine D. Plant and Giles W. Plant

31. Intraspinal Delivery of Schwann Cells for Spinal Cord Injury

Andrea J. Santamaría, Juan P. Solano, Francisco D. Benavides, and James D. Guest

Include cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts