A neural extracellular matrix-based method for in vitro hippocampal neuron culture and dopaminergic differentiation of neural stem cells

Background: The ability to recreate an optimal cellular microenvironment is critical to understand neuronal behavior and functionality in vitro. An organized neural extracellular matrix (nECM) promotes neural cell adhesion, proliferation and differentiation. Here, we expanded previous observations on the ability of nECM to support in vitro neuronal differentiation, with the following goals: (i) to recreate complex neuronal networks of embryonic rat hippocampal cells, and (ii) to achieve improved levels of dopaminergic differentiation of subventricular zone (SVZ) neural progenitor cells. Methods: Hippocampal cells from E18 rat embryos were seeded on PLL- and nECM-coated substrates. Neurosphere cultures were prepared from the SVZ of P4-P7 rat pups, and differentiation of neurospheres assayed on PLL- and nECM-coated substrates. Results: When seeded on nECM-coated substrates, both hippocampal cells and SVZ progenitor cells showed neural expression patterns that were similar to their poly-L-lysine-seeded counterparts. However, nECM-based cultures of both hippocampal neurons and SVZ progenitor cells could be maintained for longer times as compared to poly-L-lysine-based cultures. As a result, nECM-based cultures gave rise to a more branched neurite arborization of hippocampal neurons. Interestingly, the prolonged differentiation time of SVZ progenitor cells in nECM allowed us to obtain a purer population of dopaminergic neurons. Conclusions: We conclude that nECM-based coating is an efficient substrate to culture neural cells at different stages of differentiation. In addition, neural ECM-coated substrates increased neuronal survival and neuronal differentiation efficiency as compared to cationic polymers such as poly-L-lysine.

Museos y parques naturales: comunidades locales, administraciones públicas y patrimonialización de la cultura y la naturaleza

Cap. 1. Patrimonialización cultural y natural: un proceso, múltiples aproximaciones. Cap. 2. Comment un musée de ville peut-il être au service des citoyens? Un parcours et quelques pistes d’action. Jean-François Leclerc. Cap. 3. Los museos comunitarios de Kuna Yala y la memoria histórica. Anelio Merry López Cap. 4. Turismo y museos en la ciudad de Valencia. Javier Martí. Cap. 5. La Red de Museos Etnográficos de Asturias: proyecto y realidad. Juaco López Álvarez. Cap. 6. Culturas campesinas y conservación del patrimonio natur-rural. Jaime Izquierdo. Cap. 7. Faire et savoir faire un « territoire patrimonial » : Parc naturel régional du Haut-Jura (France). Olivier Givre. Cap. 8. Espacios naturales y especies salvajes. La construcción de la naturaleza como patrimonio en el Pallars Sobirà, Pirineo catalán. Oriol Beltran e Ismael Vaccaro. Cap. 9. L’histoire au cœur de la cité : l’exemple du laboratoire d’histoire et de patrimoine de Montréal. Joanne Burgess.