title: Long-term molecular and cellular stability of human neural stem cell lines.
abstract: Human Neural Stem Cells (hNSCs) are excellent candidates for in vitro and in vivo molecular, cellular, and developmental research, and also for ex-vivo gene transfer and cell therapy in the nervous system. However, hNSCs are mortal somatic cells, and thus invariably enter an irreversible growth arrest after a finite number of cell divisions in culture. It has been proposed that this is due to telomere shortening. Here, we show that long-term cultured (up to 4 years) v-myc perpetuated hNSC lines do preserve short but stable and homogeneous telomeres (TRF and Q-FISH determinations). hNSC lines (but not strains) express high levels of telomerase activity, which is activated by v-myc, as demonstrated here. Telomerase activity is not constitutive, becoming non-detectable after differentiation (in parallel to v-myc down-regulation). hNSC lines also maintain a stable cell cycle length, mitotic potential, differentiation and neuron generation capacity, and do not express senescence-associated beta-galactosidase over years, as studied here. These data, collectively, help to explain the immortal nature of v-myc-perpetuated hNSC lines, and to establish them as excellent research tools for basic and applied neurobiological and translational studies.
authors.affiliation: Department of Molecular Biology, Center of Molecular Biology Severo Ochoa, Autonomous University of Madrid, 28049 Madrid, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Villa
authors.forename: Beatriz
authors.initials: B
authors.lastname: Navarro-Galve
authors.forename: Carlos
authors.initials: C
authors.lastname: Bueno
authors.forename: Sonia
authors.initials: S
authors.lastname: Franco
authors.forename: María A
authors.initials: MA
authors.lastname: Blasco
authors.forename: Alberto
authors.initials: A
authors.lastname: Martinez-Serrano
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002453:Cell Cycle / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D002461:Cell Line, Transformed / Q000166:cytology / Q000201:enzymology*; D002470:Cell Survival / Q000235:genetics; D016922:Cellular Senescence / Q000235:genetics*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins; D015536:Down-Regulation / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D016259:Genes, myc / Q000235:genetics; D006801:Humans; D009474:Neurons / Q000166:cytology / Q000201:enzymology*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000201:enzymology*; D020871:RNA Stability / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D016615:Telomere / Q000201:enzymology* / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D004247:DNA; D019098:Telomerase