Prospective Identification, Isolation, and Profiling of a Telomerase-Expressing Subpopulation of Human Neural Stem Cells, using sox2 Enhancer-Directed Fluorescence-Activated Cell Sorting
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Media type:
E-Article
Title:
Prospective Identification, Isolation, and Profiling of a Telomerase-Expressing Subpopulation of Human Neural Stem Cells, using sox2 Enhancer-Directed Fluorescence-Activated Cell Sorting
Description:
<jats:p><jats:italic>Sox2</jats:italic>is expressed by neural stem and progenitor cells, and a<jats:italic>sox2</jats:italic>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<jats:italic>sox2</jats:italic>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<jats:sup>+</jats:sup>isolates were propagable for up to 1 year<jats:italic>in vitro</jats:italic>, and remained multilineage competent throughout. E/sox2:EGFP<jats:sup>+</jats:sup>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<jats:italic>notch</jats:italic>and<jats:italic>wnt</jats:italic>pathways, and identified multiple elements of each pathway that appear selective to human neural progenitors.<jats:italic>Sox2</jats:italic>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.</jats:p>