Nestin-specific green fluorescent protein expression in embryonic stem cell-derived neural precursor cells used for transplantation.
Expression of the enhanced green fluorescent protein (EGFP) under control of a thymidine kinase promoter/nestin second intron was specifically detected in nestin immunoreactive neural precursor cells after selection of murine embryonic stem (ES) cells in chemically defined medium. Allowing differentiation in vitro, the capacity of these cells to give rise to astroglia, oligodendroglia, and neurones was investigated. After intracerebral transplantation, long-lasting integration of precursor cells into the host tissue was observed, serving as a pool for successive neuronal and glial differentiation. EGFP expression by ES cell-derived neural precursor cells may be a valuable tool to optimize protocols for maintenance and expansion of these cells in vitro as well as in vivo after intracerebral transplantation. In addition, preparative fluorescence-activated cell sorting of EGFP-labeled neural precursor cells should be useful for standardization of a donor cell population for cell replacement therapies.
Andressen
C
C
Institute of Anatomy, University of Cologne, Cologne, Germany. christian.andressen@uni-koeln.de
Stöcker
E
E
Klinz
F J
FJ
Lenka
N
N
Hescheler
J
J
Fleischmann
B
B
Arnhold
S
S
Addicks
K
K
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian / Q000166:cytology*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins; D018380:Hematopoietic Stem Cell Transplantation; D007150:Immunohistochemistry; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D008164:Luminescent Proteins / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D009419:Nerve Tissue Proteins; D064231:Nestin; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors; D019076:Transgenes
D007381:Intermediate Filament Proteins; D008164:Luminescent Proteins; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; C577486:Nes protein, rat; D064231:Nestin; D049452:Green Fluorescent Proteins