IMPORTANT LEGAL NOTICE - The information on this site is subject to adisclaimerand acopyright notice
Contact   |   Search  
Cell factory - Back to the homepage Graphic element
Foreword Graphic element
Table of contents
Index by Index by Area Index by Partners Index by Keywords Gr&phic element
Volume 2

Organotypic brain slice cultures as an alternative to in vivo experimentation in the study of brain repair mechanisms (ORCA)



Cell factory area


EU Contribution

2,590,368 Euro


36 months



Starting date

01 January 2002

animal experimentation alternatives
neural repair protocols
stem cell application

This project is aimed at developing in vitro alternatives to animal models for the study of neurodegeneration and repair mechanisms after stroke and traumatic brain injury. The project is mainly concerned with the use and application of organotypic cultures of rat hippocampus in these studies. Organotypic cultures are grown as whole slices of brain tissue and retain many of the characteristics of the orignal brain tissue and therefore respond in a more realistic manner to brain insults such as ischaemia or trauma. The development of these models takes place in the context of an academic and industrial collaboration as four university laboratories and three industrial laboratories are participating in the consortium.


The objectives of the project are to develop new models and a common set of standardized protocols and tools with a particular emphasis on the development of strategies to study neural repair in these cultures.

A set of common tools will be developed which include the development of organotypic cultures from adolescent and adult donors, transgenic mice for real-time imaging strategies for visualizing neuronal and glial reactions to injury, techniques for assessment of anatomical and functional recovery following injury, and an assessment of the potential role for endogenous precursor cells and stem cells in post-injury repair processes. In addition two new models of traumatic brain injury will be developed to study focal (transection) and diffuse (shear) injury in organotypic brain slice cultures (slice cultures).


The workplan is structured as six work packages (WP), with three work packages specifically addressing the objectives of developing and validating new, alternative in vitro models for stroke and traumatic brain lesions, while the other three work packages are tool-oriented, providing tools for the extension of organotypic culture technologies to study repair processes. In addition existing models of ischaemia based on oxygen glucose withdrawal will be studied. Two new models of traumatic brain injury are generated in this project. The first of these involves generating and characterising a new model of focal injury based on transecting specific neuronal pathways in the slice cultures. Both histological and functional characterisation will be carried out to determine both anatomical and functional outcome measures. Stretching cultures that have been grown on elastic silicone substrates will generate a second model of axonal shear injury. This model is aimed at replicating damage seen with diffuse axonal injury and will also use both morphological and functional outcome measures. Currently it is only possible to generate slice cultures from quite young >P10 animals and it would be an advantage to be able to use older donors. Two of the groups are combining their expertise to tackle the question of why it is technically difficult to generate cultures from older donors. Transgenic mice will be generated in this project that have fluorescence reporter systems knocked-in that will allow visualization of degeneration and regeneration in organotypic cultures. This part of the project will also result in the development of transfection protocols to enable the study of novel gene function. The use of stem cells in the enhancement of repair processes will also be studies in the injury models. We will study the regulation of local neurogenesis in organotypic cultures from donors of different ages and at different stages of culture maturation, as well as examining the incorporation of normal and modified exogenous precursor cells.


The project will generate technology that extends current organotypic model technology to encompass repair mechanisms in addition to neurodegenerative mechanisms and to extend models to include traumatic injury in addition to stroke. Specifically we intend to generate two new models of traumatic injury, one focal, one diffuse. Protocols for the generation of organotypic cultures from older rats. Fluorescent transgenic mice to study degeneration and repair mechanisms in vitro.

An evaluation of the potential for endogenous neurogenesis and stem cell application to repair processes after in vitro stroke and trauma.

Dr Lars Sundström
Clinical Neurosciences
Biomedical Sciences Building
Bassett Crescent East, Southampton
S016 7PX Hants, United Kingdom
Tel: +44-23-8059 7804
Fax: +44-23-8059 7809
Prof. Dr Jens Zimmer
Head of Institute of Medical Biology
Anatomy and Neurobiology
University of Southern Denmark
Odense University
Winsloewparken 21
5000 Odense C, Denmark
Tel: +45-6550 3800/3801
Fax: +45-6590 6321

Prof. Klaus G. Reymann
Project Group Neuropharmacology
Leibniz-Institute for Neurobiology
P.O. Box 1860
Brenneckstrasse 6
39008 Magdeburg, Germany
Tel: +49-391-6263 437
Fax: +49-391-6263 438

Dr Chris Benham
Neurology Centre of Excellence for Drug Discovery
New Frontiers Science Park (North)
Third Avenue, Harlow
CM19 5AW Essex, United Kingdom
Tel: +44-1279-622 558
Fax: +44-1279-622 348

Dr Frank Striggow
Keyneurotek AG
Leipziger Strasse 44
39120 Magdeburg, Germany
Tel: +49-391-6117 200
Fax: +49-391-6117 201

Dr Jens Noraberg
Managing Director
Neuroscreen APS
c/o Anatomy and Neurobiology
University of Southern Denmark
Odense University
Winsloewparken 21
5000 Odense C, Denmark
Tel: +45-6550 3812/3810
Fax: +45-6590 6321

Dr N.A. Jensen
Laboratory of Experimental Molecular
Genetics, IMBG 6.5
The Panum Institute
Copenhagen University
Blegdamsvej 3
Copenhagen N-2200, Denmark
Tel: +45-3532 7722
Fax: +45-3532 7701