Prospective identification, isolation, and profiling of a telomerase-expressing subpopulation of human neural stem cells, using sox2 enhancer-directed fluorescence-activated cell sorting.
Sox2 is expressed by neural stem and progenitor cells, and a sox2 enhancer identifies these cells in the forebrains of both fetal and adult transgenic mouse reporters. We found that an adenovirus encoding EGFP placed under the regulatory control of a 0.4 kb sox2 core enhancer selectively identified multipotential and self-renewing neural progenitor cells in dissociates of human fetal forebrain. Upon EGFP-based fluorescence-activated cell sorting (FACS), the E/sox2:EGFP(+) isolates were propagable for up to 1 year in vitro, and remained multilineage competent throughout. E/sox2:EGFP(+) cells expressed more telomerase enzymatic activity than matched E/sox2:EGFP-depleted populations, and maintained their telomeric lengths with successive passage. Gene expression analysis of E/sox2:EGFP-sorted neural progenitor cells, normalized to the unsorted forebrain dissociates from which they derived, revealed marked overexpression of genes within the notch and wnt pathways, and identified multiple elements of each pathway that appear selective to human neural progenitors. Sox2 enhancer-based FACS thus permits the prospective identification and direct isolation of a telomerase-active population of neural stem cells from the human fetal forebrain, and the elucidation of both the transcriptome and dominant signaling pathways of these critically important cells.
Center for Translational Neuromedicine and the Department of Neurology, University of Rochester Medical Center, Rochester, New York 14642, USA. Su_Wang@urmc.rochester.edu
Su
S
Wang
Devin
D
Chandler-Militello
Gang
G
Lu
Neeta S
NS
Roy
Alex
A
Zielke
Romane
R
Auvergne
Nancy
N
Stanwood
Daniel
D
Geschwind
Giovanni
G
Coppola
Silvia K
SK
Nicolis
Fraser J
FJ
Sim
Steven A
SA
Goldman
D019070:Cell Lineage / Q000235:genetics; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000145:classification / Q000166:cytology* / Q000201:enzymology; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005333:Fetus; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D058953:Neural Stem Cells / Q000145:classification / Q000166:cytology* / Q000201:enzymology; D011446:Prospective Studies; D055748:SOXB1 Transcription Factors / Q000235:genetics*; D019098:Telomerase / Q000096:biosynthesis* / Q000235:genetics
C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D019098:Telomerase