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Neurospheres (NSs) are clonal cellular aggregates composed of neural stem cells and progenitors. A comprehensive description of their proliferation and differentiation regulation is an essential prerequisite for their use in biotherapies. Cytokines are essential molecules regulating cell precursor fate. Using a gene‐array strategy, we conducted a descriptive and functional analysis of endogenous cytokines and receptors expressed by spinal cord–derived NSs during their growth or their differentiation into neuronal and glial cells. NSs were found to express approximately 100 receptor subunits and cytokine/secreted developmental factors. Several angiogenic factors and receptors that could mediate neural precursor cell–endothelial cell relationships were detected. Among them, receptor B for endothelins was highly expressed, and endothelins were found to increase NS growth. In contrast, NSs express receptors for ciliary neurotrophic factor (CNTF), bone morphogenetic protein (BMP), interferon (IFN)‐γ, or tumor necrosis factor (TNF)‐α, which, when added in the growth phase, led to a dramatic growth reduction followed by a reduction or a loss of oligodendrocyte formation on differentiation. In addition, NSs synthesize fibroblast growth factor 2/epidermal growth factor (FGF2/EGF)–regulated endogenous cytokines that participate in their growth and differentiation. Notably, BMP‐7 and CNTF were expressed during expansion, but upon differentiation there was a remarkable switch from BMP‐7 to BMP‐4 and ‐6 and a sharp increase of CNTF. Reintroduction of growth factors reverses the BMP expression profile, indicating growth factor‐BMP cross‐regulations. The role of endogenous CNTF was investigated by deriving NSs from CNTF knockout mice. These NSs have an increased growth rate associated with reduction of apoptosis and generate astrocytes with a reduced glial fibulary acidic protein (GFAP) content. These results demonstrate the combined role of endogenous and exogenous cytokines in neural precursor cell growth and differentiation.

stem cells

Exogenous and Fibroblast Growth Factor 2/Epidermal Growth Factor–Regulated Endogenous Cytokines Regulate Neural Precursor Cell Growth and Differentiation