Regulation of Neuronal Differentiation in Human CNS Stem Cell Progeny by Leukemia Inhibitory Factor

Abstract

The generation of diverse types of neural cells during development occurs through the progressive restriction of the fate potential of neuroepithelial progenitor cells. This process is controlled by factors intrinsic and extrinsic to the cell. While the effect of extrinsic cues on multipotent stem cells of the murine central nervous system (CNS) is becoming clearer, little is known of neural stem cells of human origin. We sought to establish the roles played by two cytokines, leukemia inhibitory (LIF) and ciliary neurotrophic factor (CNTF), and by nerve growth factor (NGF) and platelet-derived growth factor (PDGF) in regulating neuronal and astroglial differentiation in cultured embryonic diencephalic human stem cells. While NGF did not influence either neuronal or glial formation, PDGF surprisingly decreased the percentage of stem cell-generated neurons, an effect opposite to that observed in murine progenitors. Furthermore, while we confirmed the known ability of LIF and CNTF to support astroglial differentiation, we also observed that, in contrast with their murine counterparts, the fraction of CNS stem cell-generated neurons in human cultures was enhanced twofold in the presence of both cytokines. These findings highlight important differences between humans and rodents in regard to the way epigenetic cues regulate the function of neural stem cells.

Isolation and Characterization of Tumorigenic, Stem-like Neural Precursors from Human Glioblastoma

“…Early passage (passages 10 -15) clonally derived glioblastoma multiforme cells underwent terminal differentiation upon removal of mitogens (34,36) and the subsequent addition of leukemia inhibitory factor to the medium, as shown previously for human fetal neural stem cells (24). Similar to normal neural stem cells (34), all of the glioblastoma multiforme clonal lines differentiated into GFAP-positive astrocytelike cells ( Fig.…”

“…To assess for multipotency, cells were plated at a density of 2.5 ϫ 10 4 cells/cm 2 onto Matrigel-coated glass coverslips (12 mm diameter) in the presence of leukemia inhibitory factor (10 ng/mL; ref. 24) for 7 to 10 days. Multiple immunofluorescence assays for neural antigens were performed as described by Gritti et al (21) and Galli et al (23).…”

Isolation and Characterization of Neural Stem Cells from the Adult Human Olfactory Bulb

“…Neural stem cells have been found in cultures of embryonic and adult murine brain, as well as embryonic human brain diencephalon, after epigenetic stimulation [13][14][15][16].…”

“…Neural stem cells with these characteristics have been found in cultures of embryonic and adult murine brain [10][11][12][13][14], as well as in human embryonic brain explants after epigenetic stimulation [15,16]. The SVZ is an intensive proliferative zone and the progeny of neural stem cells can either die or give rise to "neural progenitors" that migrate to the olfactory bulb [17].…”

Human neural progenitor cells display limited cytotoxicity and increased oligodendrogenesis during inflammation

“…[9][10][11][12][13] In order to evaluate the effect of acute stimulation with inflammatory cytokines on NPCs, we exposed neurospheres at early passages to TNF-a, IL-1b and IFN-g and analyzed the expression of 94 apoptosis-related mRNA by a custom RT-PCR array (Microfluidic Card, Applied Biosystems). Interestingly, we observed that in the presence of proinflammatory cytokines nestin-positive primitive neural cells express considerable amounts of TNF-a, TL1A and TRAIL mRNA, while showing no detectable CD95L (data not shown).…”