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J Neurosci Res. 2009 Jul;87(9):1986-96. doi: 10.1002/jnr.22041.

BDNF and extracellular matrix regulate differentiation of mice neurosphere-derived cells into a GABAergic neuronal phenotype.

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Center for Neuroscience and Cell Biology, Faculty of Medicine, University of Coimbra, Coimbra, Portugal.


Differentiation of neurosphere-derived cells is regulated by extracellular cues, namely, growth factors and proteins of the extracellular matrix (ECM). In this study we analyzed the influence of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), retinoic acid plus potassium chloride (RA-KCl), and the nonsynthetic ECMs laminin (LN) and fibronectin (FN) versus the synthetic adhesion substrate poly-L-lysine (PLL) in the in vitro differentiation of postnatal neurosphere cells. BDNF increased the number of differentiated neurons and decreased the number of neuronal precursors (nestin-positive cells) compared with NGF or RA-KCl. Moreover, cells treated with BDNF plus B27 supplement acquired a gamma-aminobutyric acid (GABA)-ergic phenotype and showed increased survival. No significant differences were found in the number of differentiated neurons in the presence of the ECMs alone. Nevertheless, FN or PLL in combination with BDNF promoted the acquisition of a GABAergic phenotype. The results obtained in this study highlight the importance of growth factors and ECM proteins for the potential of neurosphere cells to differentiate into neurons.

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