Article

Stem Cell Reviews and Reports

, Volume 8, Issue 4, pp 1178-1187

Generation of Schwann Cell-Derived Multipotent Neurospheres Isolated from Intact Sciatic Nerve

  • Ina MartinAffiliated withMolecular Neurobiology, Department of Cell Biology Faculty of Biology, University of Bielefeld
  • , The Duy NguyenAffiliated withDepartment of Cell Biology, Faculty of Biology, University of Bielefeld
  • , Vivien KrellAffiliated withDepartment of Cell Biology, Faculty of Biology, University of Bielefeld
  • , Johannes F. W. GreinerAffiliated withDepartment of Cell Biology, Faculty of Biology, University of Bielefeld
  • , Janine MüllerAffiliated withMolecular Neurobiology, Department of Cell Biology Faculty of Biology, University of Bielefeld
  • , Stefan HauserAffiliated withMolecular Neurobiology, Department of Cell Biology Faculty of Biology, University of Bielefeld
  • , Peter HeimannAffiliated withDepartment of Cell Biology, Faculty of Biology, University of Bielefeld
  • , Darius WideraAffiliated withDepartment of Cell Biology, Faculty of Biology, University of Bielefeld Email author 

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Abstract

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.

Keywords

Schwann cells Multipotency Neural crest-derived stem cells Neurosphere Cellular reprogramming