Human progenitor cells isolated from the developing cortex undergo decreased neurogenesis and eventual senescence following expansion in vitro.
Isolation of a true self-renewing stem cell from the human brain would be of great interest as a reliable source of neural tissue. Here, we report that human fetal cortical cells grown in epidermal growth factor expressed low levels of telomerase and telomeres in these cultures shortened over time leading to growth arrest after 30 weeks. Following leukemia inhibitory factor (LIF) supplementation, growth rates and telomerase expression increased. This was best demonstrated following cell cycle synchronization and staining for telomerase using immunocytochemistry. This increase in activity resulted in the maintenance of telomeres at approximately 7 kb for more than 60 weeks in vitro. However, all cultures displayed a lack of oligodendrotye production, decreases in neurogenesis over time and underwent replicative senescence associated with increased expression of p21 before 70 weeks in vitro. Thus, under our culture conditions, these cells are not stable, multipotent, telomerase expressing self-renewing stem cells. They may be more accurately described as human neural progenitor cells (hNPC) with limited lifespan and bi-potent potential (neurons/astrocytes). Interestingly, hNPC follow a course of proliferation, neuronal production and growth arrest similar to that seen during expansion and development of the human cortex, thus providing a possible model neural system. Furthermore, due to their high expansion potential and lack of tumorogenicity, these cells remain a unique and safe source of tissue for clinical transplantation.
Wright
Lynda S
LS
Waisman Center and Departments of Neurology and Anatomy University of Wisconsin-Madison, WI 53705-2280, USA.
Prowse
Karen R
KR
Wallace
Kyle
K
Linskens
Maarten H K
MH
Svendsen
Clive N
CN
D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D019070:Cell Lineage; D002478:Cells, Cultured; D016922:Cellular Senescence; D002540:Cerebral Cortex / Q000166:cytology* / Q000196:embryology; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005260:Female; D006801:Humans; D015850:Interleukin-6 / Q000494:pharmacology; D053605:Leukemia Inhibitory Factor; D009474:Neurons / Q000166:cytology*; D011247:Pregnancy; D013234:Stem Cells / Q000166:cytology*; D019098:Telomerase / Q000378:metabolism
D015850:Interleukin-6; C507837:LIF protein, human; D053605:Leukemia Inhibitory Factor; D004815:Epidermal Growth Factor; D019098:Telomerase