Telomerase immortalization of neuronally restricted progenitor cells derived from the human fetal spinal cord.
Lineage-restricted progenitors of the central nervous system (CNS) are not readily expandable because their mitotic competence is limited. Here we used retroviral overexpression of human telomerase reverse transcriptase (hTERT) to immortalize progenitors from human fetal spinal cord. The hTERT-immortalized cells divided in basic fibroblast growth factor (bFGF) expressed high telomerase activity, and gave rise to phenotypically restricted subpopulations of either glia or neurons. The latter included a prototypic line, hSC11V-TERT, that gave rise only to neurons. These included both chx10(+) interneurons and Islet1(+)/Hb9(+)/ChAT(+) motor neurons; the latter were recognized by green fluorescent protein (GFP) driven by the Hb9 enhancer. The neurons were postmitotic and achieved electrophysiologic competence. Upon xenograft to both fetal rat brain and injured adult spinal cord, they matured as neurons and survived for 6 months, with no evident tumorigenesis. The cells have survived >168 doublings in vitro, with karyotypic normalcy and without replicative senescence. hTERT overexpression thus permits the generation of progenitor lines able to give rise to phenotypically restricted neurons.
Department of Neurology and Neuroscience, Cornell University Medical Center, New York, New York 10021, USA.
Neeta S
NS
Roy
Takahiro
T
Nakano
H Michael
HM
Keyoung
Martha
M
Windrem
William K
WK
Rashbaum
M Lita
ML
Alonso
Jian
J
Kang
Weiguo
W
Peng
Melissa K
MK
Carpenter
Jane
J
Lin
Maiken
M
Nedergaard
Steven A
SA
Goldman
D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology; D004268:DNA-Binding Proteins; D024861:Genetic Enhancement / Q000379:methods; D006801:Humans; D009422:Nervous System Diseases / Q000601:surgery; D009473:Neuronal Plasticity / Q000502:physiology; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D012191:Retroviridae Proteins / Q000235:genetics / Q000378:metabolism; D013116:Spinal Cord / Q000166:cytology / Q000196:embryology / Q000502:physiology; D033581:Stem Cell Transplantation / Q000379:methods; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology*; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D023822:Tissue Engineering / Q000379:methods*
D004268:DNA-Binding Proteins; D012191:Retroviridae Proteins; C509186:TERT protein, human; D019098:Telomerase