Morphodynamic aspects of neural stem cell differentiation

WILSON C; REMEDI MM; GASTALDI L; CACERES A

Buenos Aires

Simposio; Fronteras en Biociencias 2; 2016

Instituto de Investigación en Biomedicina de Buenos Aires ? CONICET ? Instituto Partner de la Sociead Max Planck (IBioBA-MPSP)

Stem cells represent a cutting edge tool for biomedical research. In particular,neural stem cells (NSC) are especially interesting considering their potential for the development of in vitro models of neurological disorders and drug screening. However, little is known about the morphodynamic aspects behind NSCdifferentiation, which is critical considering the risks of cellular therapies, like rejection and teratoma formation.In this work we used the neurosphere assay to obtain nSC isolated from murine brains (E14) and characterize their morphological differentiation into neurons. We found that neurospheres express Nestin, β3-tubulin, MAP2 and Tau-1, suggesting that a mix of stem cells and post-mitotic neurons live inside the neurosphere. We set-up a protocol to improve both adhesion and survival of nSC after neurosphere dissociation in order to follow at the single cell level neuronal development and polarization during the first hours in culture. We found that after 3 h in culture, 67 of NSCs were positive for Nestin whereas only 33 of cells were positive for β3-tubulin. By contrast, after 72 h, 65% of cells express β3-tubulin and only 35% were Nestin+ . Interestingly, loss of Nestin immunodetection correlates with morphological changes, like breaking of the neuronal sphere, neurite outgrowth and polarization. Our data suggest that the loss of the stemness identity and first morphological changes occurred within the first 72 h in culture, suggesting that differentiation analysis should be done accordingly with this time. Further analysis will be directed to reveal neural phenotypes obtained after differentiation and the involvement of neurotrophic factors in this process.