Generation of Schwann cell-derived multipotent neurospheres isolated from intact sciatic nerve

Martin, I., Nguyen, T. D., Krell, V., Greiner, J. F. W., Müller, J., Hauser, S., Heimann, P. and Widera, D. ORCID: https://orcid.org/0000-0003-1686-130X (2012) Generation of Schwann cell-derived multipotent neurospheres isolated from intact sciatic nerve. Stem cell reviews and reports, 8 (4). pp. 1178-1187. ISSN 1558-6804 doi: 10.1007/s12015-012-9387-2

Abstract/Summary

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.