| Abstract No.: | C-B3062 |
| Country: | Canada |
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| Title: | A TISSUE-ENGINEERED MODEL TO STUDY AXONAL MIGRATION AND MYELINATION OF MOTOR NEURONS IN VITRO. |
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| Authors/Affiliations: | 2 Marie Gingras; 2 Marie-Michèle Beaulieu*; 2 Vicky Gagnon; 1 Heather D. Durham; 2 François Berthod;
1 Institut Neurologique de Montréal, QC, Canada; 2 Laboratoire d’Organogenèse Expérimentale, Québec, QC, Canada
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| Content: | Background: Amyotrophic lateral sclerosis and other motor neuron diseases are very complex as their development and progression involve many cellular processes and many cell types. To better understand and study motor neurons, in defined conditions and in a physiological environment that reflect the in vivo reality, a three-dimensional tissue-engineered in vitro model was developed by our team.
Objectives: Our aim was to study motor neurons biology in this reconstructed tissue in order to better characterize it and to further use it as a physiological in vitro model of degenerative diseases affecting motor neurons.
Methods: Purified mouse spinal cord motor neurons (1) were seeded on a collagen and chitosan sponge biomaterial populated with Schwann cells and/or fibroblasts (2). This fibroblast-populated sponge mimicked the connective tissue through which motor axons have to elongate in vivo. Tissues with motor neurons were cultured for 14, 21 and 28 days before analysis were performed. Motor neuron neurite elongation was assessed by immunofluorescence followed by microscopic observations and quantification. Myelination was assessed by immunofluorescence and transmission electronic microscopy.
Results: Our reconstructed tissue revealed itself to be a very favourable environment for the survival, neurite elongation and maturation of motor neurons. This was visualized by immunofluorescent staining of the different neurofilaments. The presence of Schwann cells in the tissue allowed myelination of the motor neuron axons. Myelin basic protein expression by Schwann cells as well as electronic microscopy observation of myelin sheaths demonstrated the abundant myelination that occurred in the tissue. Moreover, Schwann cell promoted the elongation of more motor neuron neurites in the tissue.
Discussion: This indicated that our tissue is a representative model of the in vivo physiology of motor neurons. All these characteristics make our reconstructed tissue well-suited for in vitro motor neuron studies that require a physiological three-dimensional environment.
Conclusion: Comprehension of amyotrophic lateral sclerosis and other motor neuron diseases pathogenesis will surely be greatly improved by the use of this highly promising model.
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