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2019-06-25

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Si us plau utilitzeu sempre aquest identificador per citar o enllaçar aquest document: https://hdl.handle.net/2445/152003

Generation of 3D collagen-based hydrogels to analyze axonal growth and behavior during nervous system development

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Recurs relacionat

https://doi.org/10.3791/59481

Resum

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.

Matèries

Neurones, Creixement

Matèries (anglès)

Neurons, Growth

Citació

Col·leccions

Articles publicats en revistes (Biologia Cel·lular, Fisiologia i Immunologia)

Citació

GIL FERNÁNDEZ, Vanessa and RÍO FERNÁNDEZ, José Antonio del. Generation of 3D collagen-based hydrogels to analyze axonal growth and behavior during nervous system development. JoVE. Journal of Visualized Experiments. 2019. Vol. 148, num. e59481. ISSN 1940-087X. [consulted: 7 of August of 2026]. Available at: https://hdl.handle.net/2445/152003

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