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Nat Protoc. 2012 Jan 19;7(2):268-80. doi: 10.1038/nprot.2011.445.

Analysis of axonal growth and cell migration in 3D hydrogel cultures of embryonic mouse CNS tissue.

Author information

1
Molecular and Cellular Neurobiotechnology, Institute for Bioengineering of Catalonia, Science Park of Barcelona, Barcelona, Spain.

Abstract

This protocol uses rat tail-derived type I collagen hydrogels to analyze key processes in developmental neurobiology, such as chemorepulsion and chemoattraction. The method is based on culturing small pieces of brain tissue from embryonic or early perinatal mice inside a 3D hydrogel formed by rat tail-derived type I collagen or, alternatively, by commercial Matrigel. The neural tissue is placed in the hydrogel with other brain tissue pieces or cell aggregates genetically modified to secrete a particular molecule that can generate a gradient inside the hydrogel. The present method is uncomplicated and generally reproducible, and only a few specific details need to be considered during its preparation. Moreover, the degree and behavior of axonal growth or neural migration can be observed directly using phase-contrast, fluorescence microscopy or immunocytochemical methods. This protocol can be carried out in 4 weeks.

PMID:
22262008
DOI:
10.1038/nprot.2011.445
[Indexed for MEDLINE]

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