Labeling Schwann cells with CFSE-an in vitro and in vivo study

J Neurosci Methods. 2003 May 30;125(1-2):83-91. doi: 10.1016/s0165-0270(03)00044-x.


Schwann cell (SC) transplantation is a promising strategy for axonal regeneration in the nervous system. Identifying the grafted SCs is an important aspect of this approach. The current study sought to establish a simple, reliable, fluorescent labeling method for SCs with a lipophilic molecule, 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE). Human SCs were incubated with varying concentrations of CFSE for different time periods. Based on the viability of labeled SCs and its plating efficiency, 1 min incubation with 5 microM CFSE at 37 degrees C was selected as the optimal labeling condition. Flow cytometric analysis and fluorescence microscopy demonstrated that the fluorescence of labeled SCs would fade over 4 weeks. Immunostaining for the phenotypic expression of SC markers, including S100, GFAP, P75, and MHC-I/II at 1 and 4 weeks after incubation with CFSE showed no difference between labeled and non-labeled SCs. Mixed cultures of labeled human SCs and rat SCs for 48 h were performed in triplicate and demonstrated that no leakage of dye from labeled SCs in cell culture occurred across species. A total of 14 injections of 2x10(5) labeled SCs were performed within the spinal cord at T8 and/or L1 level in 9 nude rats. The animals were euthanized at 1 (6 injections) and 4 weeks (8 injections). Grafted labeled SCs survived for at least 4 weeks, and could be easily recognized in the nude rat spinal cord without leakage of dye to surrounding cells. The SCs migrated in white and gray matter 3-6 mm away from the injection and in the central canal for up to 12 mm. These results suggest that CFSE can be used as a fluorescent tracer of human SCs for both in vitro and in vivo studies, for a period of at least 4 weeks.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Benzimidazoles / pharmacokinetics
  • Cells, Cultured
  • Coculture Techniques
  • Dose-Response Relationship, Drug
  • Female
  • Flow Cytometry
  • Fluoresceins / metabolism*
  • Fluorescent Dyes / metabolism*
  • Humans
  • Immunohistochemistry
  • Phenotype
  • Rats
  • Rats, Inbred F344
  • Rats, Nude
  • Schwann Cells / drug effects
  • Schwann Cells / metabolism*
  • Schwann Cells / transplantation
  • Spinal Cord / drug effects
  • Spinal Cord / metabolism
  • Spinal Cord / transplantation
  • Succinimides / metabolism*
  • Time Factors
  • Transplantation


  • 5-(6)-carboxyfluorescein diacetate succinimidyl ester
  • Benzimidazoles
  • Fluoresceins
  • Fluorescent Dyes
  • Succinimides
  • bisbenzimide ethoxide trihydrochloride