Thick collagen-based 3D matrices including growth factors to induce neurite outgrowth. Auteur(s): Labour M.-N., Banc A., Tourrette Audrey, Cunin Frédérique, Verdier Jean-Michel, Devoisselle Jean-Marie, Marcilhac Anne, Belamie Emmanuel (Article) Publié: Acta Biomaterialia, vol. 8 p.3302-12 (2012) Texte intégral en Openaccess : Ref HAL: hal-00747635_v1 PMID 22617741 Ref Arxiv: 1211.0301 DOI: 10.1016/j.actbio.2012.05.015 WoS: 000307625900012 Ref. & Cit.: NASA ADS Exporter : BibTex | endNote 15 Citations Résumé: Designing synthetic microenvironments for cellular investigations is a very active area of research at the crossroads of cell biology and materials science. The present work describes the design and functionalization of a three-dimensional (3D) culture support dedicated to the study of neurite outgrowth from neural cells. It is based on a dense self-assembled collagen matrix stabilized by 100-nm-wide interconnected native fibrils without chemical crosslinking. The matrices were made suitable for cell manipulation and direct observation in confocal microscopy by anchoring them to traditional glass supports with a calibrated thickness of ∼50μm. The matrix composition can be readily adapted to specific neural cell types, notably by incorporating appropriate neurotrophic growth factors. Both PC-12 and SH-SY5Y lines respond to growth factors (nerve growth factor and brain-derived neurotrophic factor, respectively) impregnated and slowly released from the support. Significant neurite outgrowth is reported for a large proportion of cells, up to 66% for PC12 and 49% for SH-SY5Y. It is also shown that both growth factors can be chemically conjugated (EDC/NHS) throughout the matrix and yield similar proportions of cells with longer neurites (61% and 52%, respectively). Finally, neurite outgrowth was observed over several tens of microns within the 3D matrix, with both diffusing and immobilized growth factors. |