The Open UniversitySkip to content
 

Engineering an integrated cellular interface in three-dimensional hydrogel cultures permits monitoring of reciprocal astrocyte and neuronal responses

East, Emma; Golding, Jon P. and Phillips, James B. (2012). Engineering an integrated cellular interface in three-dimensional hydrogel cultures permits monitoring of reciprocal astrocyte and neuronal responses. Tissue Engineering Part C: Methods, 18(7) pp. 526–536.

Full text available as:
[img]
Preview
PDF (Accepted Manuscript) - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader
Download (1578Kb)
URL: http://online.liebertpub.com/doi/abs/10.1089/ten.T...
DOI (Digital Object Identifier) Link: http://dx.doi.org/10.1089/ten.TEC.2011.0587
Google Scholar: Look up in Google Scholar

Abstract

This study reports a new type of 3D tissue model for studying interactions between cell types in collagen hydrogels. The aim was to create a 3D cell culture model containing separate cell populations in close proximity without the presence of a mechanical barrier, and demonstrate its relevance to modelling the axon growth-inhibitory cellular interfaces that develop in the central nervous system (CNS) in response to damage. This provides a powerful new tool to determine which aspects of the astroglial scar response and subsequent neuronal regeneration inhibition are determined by the presence of the other cell types. Astrocytes (CNS glia) and dissociated dorsal root ganglia (DRG; containing neurons and peripheral nervous system [PNS] glia) were seeded within collagen solution at 4°C in adjacent chambers of a stainless steel mould, using cells cultured from wild type or green fluorescent protein expressing rats, in order to track specific populations. The divider between the chambers was removed using a protocol that allowed the gels to integrate without mixing of the cell populations. Following setting of the gels, they were maintained in culture for up to 15 days. Reciprocal astrocyte and neuronal responses were monitored using confocal microscopy and 3D image analysis. At DRG:astrocyte interfaces, by 5 days there was an increase in the number of astrocytes at the interface followed by hypertrophy and increased glial fibrillary acidic protein expression at 10 and 15 days, indicative of reactive gliosis. Neurons avoided crossing DRG:astrocyte interfaces, and neuronal growth was restricted to the DRG part of the gel. By contrasct, neurons were able to grow freely across DRG:DRG interfaces, demonstrating the absence of a mechanical barrier. These results show that in a precisely controlled 3D environment, an interface between DRG and astrocyte cultures is sufficient to trigger reactive gliosis and inhibition of neuronal regeneration across the interface. Different aspects of the astrocyte response could be monitored independently, providing an insight into the formation of a glial scar. This technology has wide potential for researchers wishing to maintain and monitor interactions between adjacent cell populations in 3D culture.

Item Type: Journal Article
Copyright Holders: 2012 Mary Ann Liebert, Inc
ISSN: 1937-3392
Project Funding Details:
Funded Project NameProject IDFunding Body
Not SetNot SetThe Wellcome Trust
Keywords: spinal cord injury; nervous system; astrocyte; neuron; 3D culture; collagen gel; interface; glial scar; CNS
Academic Unit/Department: Science > Life, Health and Chemical Sciences
Science
Interdisciplinary Research Centre: Biomedical Research Network (BRN)
Related URLs:
Item ID: 32596
Depositing User: James Phillips
Date Deposited: 15 Feb 2012 11:26
Last Modified: 13 Mar 2014 22:59
URI: http://oro.open.ac.uk/id/eprint/32596
Share this page:

Actions (login may be required)

View Item
Report issue / request change

Policies | Disclaimer

© The Open University   + 44 (0)870 333 4340   general-enquiries@open.ac.uk