Isolation and characterization of neural precursor cells from the Sox1-GFP reporter mouse.
We have made use of a reporter mouse line in which enhanced green fluorescence protein (GFP) is inserted into the Sox1 locus. We show that the GFP reporter is coexpressed with the Sox1 protein as well as with other known markers for neural stem and progenitor cells, and can be used to identify and isolate these cells by fluorescence-activated cell sorting (FACS) from the developing or adult brain and from neurosphere cultures. All neurosphere-forming cells with the capacity for multipotency and self-renewal reside in the Sox1-GFP-expressing population. Thus, the Sox1-GFP reporter system is highly useful for identification, isolation and characterization of neural stem and progenitor cells, as well as for the validation of alternative means for isolating neural stem and progenitor cells. Further, transplantation experiments show that Sox1-GFP cells isolated from the foetal brain give rise to neurons and glia in vivo, and that many of the neurons display phenotypic characteristics appropriate for the developing brain region from which the Sox1-GFP precursors were derived. On the other hand, Sox1-GFP cells isolated from the adult subventricular zone or expanded neurosphere cultures gave rise almost exclusively to glial cells following transplantation. Thus, not all Sox1-GFP cells possess the same capacity for neuronal differentiation in vivo.
Barraud
Perrine
P
Wallenberg Neuroscience Center, BMC A11, Section of Neurobiology, and Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, Lund, University, SE-221 84 Lund, Sweden.
Thompson
Lachlan
L
Kirik
Deniz
D
Björklund
Anders
A
Parmar
Malin
M
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000378:metabolism; D011125:Adenomatous Polyposis Coli / Q000378:metabolism; D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000166:cytology* / Q000378:metabolism; D002452:Cell Count / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002469:Cell Separation; D002478:Cells, Cultured; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004622:Embryo, Mammalian; D005434:Flow Cytometry / Q000379:methods; D005786:Gene Expression Regulation / Q000502:physiology*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D006609:High Mobility Group Proteins / Q000378:metabolism*; D007150:Immunohistochemistry / Q000379:methods; D007381:Intermediate Filament Proteins / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008863:Microspheres; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D039842:Neural Cell Adhesion Molecule L1 / Q000378:metabolism; D009474:Neurons / Q000502:physiology*; D010320:Parvalbumins / Q000378:metabolism; D009195:Peroxidase / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D051381:Rats; D055748:SOXB1 Transcription Factors; D012794:Sialic Acids / Q000378:metabolism; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000502:physiology*; D014404:Tubulin / Q000378:metabolism
D004268:DNA-Binding Proteins; D005904:Glial Fibrillary Acidic Protein; D006609:High Mobility Group Proteins; D007381:Intermediate Filament Proteins; C491296:Msi1 protein, rat; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; C577486:Nes protein, rat; D064231:Nestin; D039842:Neural Cell Adhesion Molecule L1; D010320:Parvalbumins; D016601:RNA-Binding Proteins; D055748:SOXB1 Transcription Factors; D012794:Sialic Acids; C098693:Sox1 protein, mouse; D014404:Tubulin; C103469:polysialyl neural cell adhesion molecule; D049452:Green Fluorescent Proteins; D009195:Peroxidase; D002795:Choline O-Acetyltransferase; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; D010751:Phosphopyruvate Hydratase