Long-term molecular and cellular stability of human neural stem cell lines.
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.
Villa
Ana
A
Department of Molecular Biology, Center of Molecular Biology Severo Ochoa, Autonomous University of Madrid, 28049 Madrid, Spain.
Navarro-Galve
Beatriz
B
Bueno
Carlos
C
Franco
Sonia
S
Blasco
María A
MA
Martinez-Serrano
Alberto
A
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
D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D004247:DNA; D019098:Telomerase