Adult bone marrow-derived mononuclear cells expressing chondroitinase AC transplanted into CNS injury sites promote local brain chondroitin sulphate degradation

• Published in: Journal of neuroscience methods Vol. 171; no. 1; pp. 19 - 29
• Main Authors: Coulson-Thomas, Yvette M., Coulson-Thomas, Vivien J., Filippo, Thais R., Mortara, Renato A., da Silveira, Rafael B., Nader, Helena B., Porcionatto, Marimélia A.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15-06-2008
• Subjects: Animals; Bone Marrow Cells - metabolism; Bone Marrow Transplantation - methods; Bone marrow-derived cells; Brain Injuries - metabolism; Brain Injuries - surgery; Cell Line; Chondroitin Sulfates - metabolism; Chondroitin sulphate; Chondroitinase AC; Chondroitinases and Chondroitin Lyases - genetics; Chondroitinases and Chondroitin Lyases - metabolism; Cricetinae; Cricetulus; Female; GAP-43 Protein - metabolism; Gene Expression; Gliosarcoma; Glycosaminoglycans - metabolism; Green Fluorescent Proteins - biosynthesis; Injury; Mice; Mice, Inbred C57BL; Mice, Transgenic; Nervous system; Transfection - methods; Bone marrow-derived cells; Nervous system; Chondroitinase AC; Chondroitin sulphate; Injury
• ISSN: 0165-0270; 1872-678X
• DOI: 10.1016/j.jneumeth.2008.01.030

Injury to the CNS of vertebrates leads to the formation of a glial scar and production of inhibitory molecules, including chondroitin sulphate proteoglycans. Various studies suggest that the sugar component of the proteoglycan is responsible for the inhibitory role of these compounds in axonal regeneration. By degrading chondroitin sulphate chains with specific enzymes, denominated chondroitinases, the inhibitory capacity of these proteoglycans is decreased. Chondroitinase administration involves frequent injections of the enzyme at the lesion site which constitutes a rather invasive method. We have produced a vector containing the gene for
Flavobacterium heparinum chondroitinase AC for expression in adult bone marrow-derived cells which were then transplanted into an injury site in the CNS. The expression and secretion of active chondroitinase AC was observed
in vitro using transfected Chinese hamster ovarian and gliosarcoma cells and
in vivo by immunohistochemistry analysis which showed degraded chondroitin sulphate coinciding with the location of transfected bone marrow-derived cells. Immunolabelling of the axonal growth-associated protein GAP-43 was observed
in vivo and coincided with the location of degraded chondroitin sulphate. We propose that bone marrow-derived mononuclear cells, transfected with our construct and transplanted into CNS, could be a potential tool for studying an alternative chondroitinase AC delivery method.