Electroporation-based gene transfer for efficient transfection of neural precursor cells

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Ines Richard - , University Hospital Hamburg Eppendorf (Author)
  • Marius Ader - , University Hospital Hamburg Eppendorf, University of Hamburg (Author)
  • Vladimir Sytnyk - , University of Hamburg (Author)
  • Alexander Dityatev - , University of Hamburg (Author)
  • Gisbert Richard - , University Hospital Hamburg Eppendorf (Author)
  • Melitta Schachner - , University of Hamburg (Author)
  • Udo Bartsch - , University Hospital Hamburg Eppendorf, University of Hamburg (Author)

Abstract

Transplantation of neural precursor cells (NPCs) is a potential tool to replace dysfunctional or degenerated neuronal or glial cell types in the central nervous system. Furthermore, transplantation of genetically engineered neural precursor cells might provide a strategy to target therapeutic gene products to the diseased nervous system. Here, we describe a novel and highly efficient electroporation-based transfection protocol for mitogen-expanded mouse NPCs. Transfection of NPCs with the reporter gene enhanced green fluorescent protein (EGFP) or the neural adhesion molecule L1 revealed transfection efficacies of more than 70% as estimated by the number of EGFP-positive or L1-immunoreactive cells 1 day after transfection in vitro. The percentage of EGFP- or L1-positive cells decreased with increasing time in culture. Positive cells were detectable for up to 3 weeks after transfection. When EGFP- or L1-transfected NPCs were grafted into the retina of adult wild-type or L1-deficient mice, they differentiated into glial cells some of which expressed EGFP and L1 for up to 2 and 3 weeks, respectively, the longest post-transplantation periods investigated.

Details

Original languageEnglish
Pages (from-to)182-90
Number of pages9
JournalBrain research : an international multidisciplinary journal devoted to fundamental research in the brain sciences
Volume138
Issue number2
Publication statusPublished - 18 Aug 2005
Peer-reviewedYes
Externally publishedYes

External IDs

Scopus 23744483127
ORCID /0000-0001-9467-7677/work/161888211

Keywords

Keywords

  • Animals, CD56 Antigen/genetics, Cell Count, Cell Differentiation/genetics, Cells, Cultured, Electroporation/methods, Genes, Reporter/genetics, Green Fluorescent Proteins/genetics, Mice, Mice, Inbred C57BL, Mice, Transgenic, Neuroglia/cytology, Neurons/cytology, Stem Cell Transplantation/methods, Stem Cells/cytology, Transfection/methods, Up-Regulation/genetics