title: Thick collagen-based 3D matrices including growth factors to induce neurite outgrowth.
abstract: 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.
authors.affiliation: Ecole Pratique des Hautes Etudes, 46 rue de Lille, 75007 Paris, France.
authors.forename: M-N
authors.initials: MN
authors.lastname: Labour
authors.forename: A
authors.initials: A
authors.lastname: Banc
authors.forename: A
authors.initials: A
authors.lastname: Tourrette
authors.forename: F
authors.initials: F
authors.lastname: Cunin
authors.forename: J-M
authors.initials: JM
authors.lastname: Verdier
authors.forename: J-M
authors.initials: JM
authors.lastname: Devoisselle
authors.forename: A
authors.initials: A
authors.lastname: Marcilhac
authors.forename: E
authors.initials: E
authors.lastname: Belamie
mesh_terms: D000818:Animals; D002460:Cell Line; D003094:Collagen / Q000494:pharmacology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000494:pharmacology*; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D016501:Neurites; D051381:Rats
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D003094:Collagen