Generation of Schwann Cell-Derived Multipotent Neurospheres Isolated from Intact Sciatic Nerve
Rent the article at a discountRent now
* Final gross prices may vary according to local VAT.Get Access
Schwann cells (SCs) are the supporting cells of the peripheral nervous system and originate from the neural crest. They play a unique role in the regeneration of injured peripheral nerves and have themselves a highly unstable phenotype as demonstrated by their unexpectedly broad differentiation potential. Thus, SCs can be considered as dormant, multipotent neural crest-derived progenitors or stem cells. Upon injury they de-differentiate via cellular reprogramming, re-enter the cell cycle and participate in the regeneration of the nerve. Here we describe a protocol for efficient generation of neurospheres from intact adult rat and murine sciatic nerve without the need of experimental in vivo pre-degeneration of the nerve prior to Schwann cell isolation. After isolation and removal of the connective tissue, the nerves are initially plated on poly-D-lysine coated cell culture plates followed by migration of the cells up to 80 % confluence and a subsequent switch to serum-free medium leading to formation of multipotent neurospheres. In this context, migration of SCs from the isolated nerve, followed by serum-free cultivation of isolated SCs as neurospheres mimics the injury and reprograms fully differentiated SCs into a multipotent, neural crest-derived stem cell phenotype. This protocol allows reproducible generation of multipotent Schwann cell-derived neurospheres from sciatic nerve through cellular reprogramming by culture, potentially marking a starting point for future detailed investigations of the de-differentiation process.
- Kaltschmidt, B., Kaltschmidt, C., & Widera, D. (2011). Adult craniofacial stem cells: sources and relation to the neural crest. Stem Cell Reviews. doi:10.1007/s12015-011-9340-9.
- Schwann, T. (1839). Mikroskopische Untersuchungen über die Uebereinstimmung in der Struktur und dem Wachsthum der Thiere und Pflanzen. Berlin: Verlag der Sanderschen Buchhandlung.
- Bunge, R. P. (1993). Expanding roles for the Schwann cell: ensheathment, myelination, trophism and regeneration. Current Opinion in Neurobiology, 3, 805–809. CrossRef
- Fawcett, J. W., & Keynes, R. J. (1990). Peripheral nerve regeneration. Annual Review of Neuroscience, 13, 43–60. CrossRef
- Mirsky, R., Jessen, K. R., Brennan, A., et al. (2002). Schwann cells as regulators of nerve development. Journal of Physiology, Paris, 96, 17–24. CrossRef
- Sherman, L., Stocker, K. M., Morrison, R., & Ciment, G. (1993). Basic fibroblast growth factor (bFGF) acts intracellularly to cause the transdifferentiation of avian neural crest-derived Schwann cell precursors into melanocytes. Development, 118, 1313–1326.
- Dupin, E., Real, C., Glavieux-Pardanaud, C., Vaigot, P., & Le Douarin, N. M. (2003). Reversal of developmental restrictions in neural crest lineages: transition from Schwann cells to glial-melanocytic precursors in vitro. Proceedings of the National Academy of Sciences of the United States of America, 100, 5229–5233. CrossRef
- Real, C., Glavieux-Pardanaud, C., Vaigot, P., Le-Douarin, N., & Dupin, E. (2005). The instability of the neural crest phenotypes: Schwann cells can differentiate into myofibroblasts. The International Journal of Developmental Biology, 49, 151–159. CrossRef
- Binder, E., Rukavina, M., Hassani, H., et al. (2011). Peripheral nervous system progenitors can be reprogrammed to produce myelinating oligodendrocytes and repair brain lesions. The Journal of Neuroscience: The Official Journal of The Society for Neuroscience, 31, 6379–6391. CrossRef
- Widera, D., Heimann, P., Zander, C., et al. (2011). Schwann cells can be reprogrammed to multipotency by culture. Stem Cells and Development, 20, 2053–2064. CrossRef
- Morrissey, T. K., Kleitman, N., & Bunge, R. P. (1991). Isolation and functional characterization of Schwann cells derived from adult peripheral nerve. The Journal of Neuroscience: The Official Journal of The Society for Neuroscience, 11, 2433–2442.
- Takagi, T., Ishii, K., Shibata, S., et al. (2011). Schwann-spheres derived from injured peripheral nerves in adult mice–their in vitro characterization and therapeutic potential. PloS One, 6, e21497. CrossRef
- Popovic, M., Sketelj, J., & Bresjanac, M. (1996). Changes of Schwann cell antigenic profile after peripheral nerve injury. Pflugers Archiv: European Journal of Physiology, 431, R287–R288. CrossRef
- Rutkowski, J. L., Tennekoon, G. I., & McGillicuddy, J. E. (1992). Selective culture of mitotically active human Schwann cells from adult sural nerves. Annals of Neurology, 31, 580–586. CrossRef
- Keilhoff, G., Fansa, H., Schneider, W., & Wolf, G. (1999). In vivo predegeneration of peripheral nerves: an effective technique to obtain activated Schwann cells for nerve conduits. Journal of Neuroscience Methods, 89, 17–24. CrossRef
- Verdu, E., Rodriguez, F. J., Gudino-Cabrera, G., Nieto-Sampedro, M., & Navarro, X. (2000). Expansion of adult Schwann cells from mouse predegenerated peripheral nerves. Journal of Neuroscience Methods, 99, 111–117. CrossRef
- de la Fuente, I., Alcalde, I., Gamboa, O. L., Garrosa, M., & Gayoso, M. J. (2012). A method for obtaining Schwann cell cultures from adult rabbit nerve based on “in vitro” pre-degeneration and neuregulin treatment. Histology and Histopathology, 27, 95–102.
- Simm, A., Bertsch, G., Frank, H., Zimmermann, U., & Hoppe, J. (1997). Cell death of AKR-2B fibroblasts after serum removal: a process between apoptosis and necrosis. Journal of Cell Science, 110(Pt 7), 819–828.
- Russel, S. A. (2001). Molecular cloning: a laboratory manual. 1.
- Chen, Y., Stevens, B., Chang, J., Milbrandt, J., Barres, B. A., & Hell, J. W. (2008). NS21: re-defined and modified supplement B27 for neuronal cultures. Journal of Neuroscience Methods, 171, 239–247. CrossRef
- Generation of Schwann Cell-Derived Multipotent Neurospheres Isolated from Intact Sciatic Nerve
Stem Cell Reviews and Reports
Volume 8, Issue 4 , pp 1178-1187
- Cover Date
- Print ISSN
- Online ISSN
- Humana Press Inc
- Additional Links
- Schwann cells
- Neural crest-derived stem cells
- Cellular reprogramming
- Industry Sectors
- Author Affiliations
- 1. Molecular Neurobiology, Department of Cell Biology Faculty of Biology, University of Bielefeld, Bielefeld, Germany
- 2. Department of Cell Biology, Faculty of Biology, University of Bielefeld, Bielefeld, Germany