Abstract
This chapter emphasizes detailed protocols for the effective establishment of highly enriched human Schwann cell cultures and their characterization via immunostaining. The Schwann cells are isolated from immediately dissociated fascicle tissue and expanded prior to purification. Two purification methods are described that use either fluorescence-activated cell sorting for the Schwann cell marker TNR16 (p75NTR) or a less-manipulative two-step enrichment exploiting the differential adhesion properties of Schwann cells and fibroblasts, which is especially useful for low Schwann cell numbers. In addition, a method to determine Schwann cell purity via stained cytospin slides is introduced. Together with an immunofluorescence staining procedure for the combined analysis of extra- and intracellular markers, this chapter provides a solid basis to study human primary Schwann cells.
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Acknowledgments
We wish to thank Dieter Printz (FACS core unit, Children’s Cancer Research Institute, Vienna, Austria) for cell sorting and Prof. Dr. Reinhard Windhager (Department of Orthopedic Surgery, Medical University of Vienna, Austria), Dr. Hugo B. Kitzinger and Prof. Dr. Chieh-Han Tzou (Division of Plastic and Reconstructive Surgery, Medical University of Vienna, Austria), and their patients for providing human nerve tissue samples. The research leading to these results has received funding from the St. Anna Kinderkrebsforschung (Vienna, Austria) and an FFG (TisQuant, EraSME, by the Austrian Research Promotion Agency) grant to Peter F. Ambros.
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Weiss, T., Taschner-Mandl, S., Ambros, P.F., Ambros, I.M. (2018). Detailed Protocols for the Isolation, Culture, Enrichment and Immunostaining of Primary Human Schwann Cells. In: Monje, P., Kim, H. (eds) Schwann Cells. Methods in Molecular Biology, vol 1739. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7649-2_5
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DOI: https://doi.org/10.1007/978-1-4939-7649-2_5
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