Generation of Schwann Cell-Derived Multipotent Neurospheres Isolated from Intact Sciatic Nerve
- 542 Downloads
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.
KeywordsSchwann cells Multipotency Neural crest-derived stem cells Neurosphere Cellular reprogramming
This study was supported by a FiF grant of the Bielefeld University to DW.
Conflicts of interest
The authors declare no potential conflicts of interest.
- 1.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.
- 2.Schwann, T. (1839). Mikroskopische Untersuchungen über die Uebereinstimmung in der Struktur und dem Wachsthum der Thiere und Pflanzen. Berlin: Verlag der Sanderschen Buchhandlung.Google Scholar
- 7.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.PubMedCrossRefGoogle Scholar
- 11.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.Google Scholar
- 19.Russel, S. A. (2001). Molecular cloning: a laboratory manual. 1.Google Scholar