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Title: Generation of 3D collagen-based hydrogels to analyze axonal growth and behavior during nervous system development
Author: Gil Fernández, Vanessa
Río Fernández, José Antonio del
Keywords: Neurones
Issue Date: 25-Jun-2019
Publisher: JoVE
Abstract: This protocol uses natural type I collagen to generate three-dimensional (3-D) hydrogel for monitoring and analyzing the axonal growth. The protocol is centered on culturing small pieces of embryonic or early postnatal rodent brains inside a 3-D hydrogel formed by the rat tail tendon-derived type I collagen with specific porosity. Tissue pieces are cultured inside the hydrogel and confronted to specific brain fragments or genetically-modified cell aggregates to produce and secrete molecules suitable for creating a gradient inside the porous matrix. The steps of this protocol are simple and reproducible but include critical steps to be considered carefully during its development. Moreover, the behavior of growing axons can be monitored and analyzed directly using a phase-contrast microscope or mono/multiphoton fluorescence microscope after fixation by immunocytochemical methods.
Note: Reproducció del document publicat a:
It is part of: JoVE. Journal of Visualized Experiments, 2019, vol. 148, p. e59481
Related resource:
ISSN: 1940-087X
Appears in Collections:Articles publicats en revistes (Biologia Cel·lular, Fisiologia i Immunologia)

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