Electroporation-based gene transfer for efficient transfection of neural precursor cells
Standard
Electroporation-based gene transfer for efficient transfection of neural precursor cells. / Richard, Ines; Ader, Marius; Sytnyk, Vladimir; Dityatev, Alexander; Richard, Gisbert; Schachner, Melitta; Bartsch, Udo.
In: MOL BRAIN RES, Vol. 138, No. 2, 18.08.2005, p. 182-90.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
Harvard
APA
Vancouver
Bibtex
}
RIS
TY - JOUR
T1 - Electroporation-based gene transfer for efficient transfection of neural precursor cells
AU - Richard, Ines
AU - Ader, Marius
AU - Sytnyk, Vladimir
AU - Dityatev, Alexander
AU - Richard, Gisbert
AU - Schachner, Melitta
AU - Bartsch, Udo
PY - 2005/8/18
Y1 - 2005/8/18
N2 - 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.
AB - 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.
KW - Animals
KW - Antigens, CD56
KW - Cell Count
KW - Cell Differentiation
KW - Cells, Cultured
KW - Electroporation
KW - Genes, Reporter
KW - Green Fluorescent Proteins
KW - Mice
KW - Mice, Inbred C57BL
KW - Mice, Transgenic
KW - Neuroglia
KW - Neurons
KW - Stem Cell Transplantation
KW - Stem Cells
KW - Transfection
KW - Up-Regulation
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1016/j.molbrainres.2005.04.010
DO - 10.1016/j.molbrainres.2005.04.010
M3 - SCORING: Journal article
C2 - 15908040
VL - 138
SP - 182
EP - 190
IS - 2
ER -