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

Xiuming Li, Hector Dancausse, Israel Grijalva, Maria Oliveira, Allan D Levi

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

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 μM CFSE at 37°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 2×105 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.

Original languageEnglish
Pages (from-to)83-91
Number of pages9
JournalJournal of Neuroscience Methods
Volume125
Issue number1-2
DOIs
StatePublished - May 30 2003

Fingerprint

Schwann Cells
Nude Rats
Injections
Spinal Cord
Coloring Agents
Cell Transplantation
Fluorescence Microscopy
Nervous System
In Vitro Techniques
5-(6)-carboxyfluorescein diacetate succinimidyl ester
Regeneration
Cell Culture Techniques
Fluorescence

Keywords

  • CFSE
  • Fluorescence
  • Labeling
  • Nude rats
  • Schwann cells
  • Spinal cord
  • Transplantation

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Labeling Schwann cells with CFSE - An in vitro and in vivo study. / Li, Xiuming; Dancausse, Hector; Grijalva, Israel; Oliveira, Maria; Levi, Allan D.

In: Journal of Neuroscience Methods, Vol. 125, No. 1-2, 30.05.2003, p. 83-91.

Research output: Contribution to journalArticle

Li, Xiuming ; Dancausse, Hector ; Grijalva, Israel ; Oliveira, Maria ; Levi, Allan D. / Labeling Schwann cells with CFSE - An in vitro and in vivo study. In: Journal of Neuroscience Methods. 2003 ; Vol. 125, No. 1-2. pp. 83-91.
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