An in vitro model of the inhibition of axon growth in the lesion scar formed after central nervous system injury.
After central nervous system (CNS) injury, meningeal fibroblasts migrate in the lesion center to form a fibrotic scar which is surrounded by end feet of reactive astrocytes. The fibrotic scar expresses various axonal growth-inhibitory molecules and creates a major impediment for axonal regeneration. We developed an in vitro model of the scar using coculture of cerebral astrocytes and meningeal fibroblasts by adding transforming growth factor-beta1 (TGF-beta1), a potent fibrogenic factor. Addition of TGF-beta1 to this coculture resulted in enhanced proliferation of fibroblasts and the formation of cell clusters which consisted of fibroblasts inside and surrounded by astrocytes. The cell cluster in culture densely accumulated the extracellular matrix molecules and axonal growth-inhibitory molecules similar to the fibrotic scar, and remarkably inhibited the neurite outgrowth of cerebellar neurons. Therefore, this culture system can be available to analyze the inhibitory property in the lesion site of CNS.
Kimura-Kuroda
Junko
J
Department of Developmental Morphology, Tokyo Metropolitan Institute for Neuroscience, Tokyo, Japan.
Teng
Xichuan
X
Komuta
Yukari
Y
Yoshioka
Nozomu
N
Sango
Kazunori
K
Kawamura
Koki
K
Raisman
Geoffrey
G
Kawano
Hitoshi
H
D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D000941:Antigens / Q000378:metabolism; D001253:Astrocytes / Q000187:drug effects / Q000502:physiology; D001369:Axons / Q000502:physiology*; D001973:Bromodeoxyuridine / Q000378:metabolism; D002452:Cell Count; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology; D002540:Cerebral Cortex / Q000166:cytology; D002809:Chondroitin Sulfates / Q000378:metabolism; D018920:Coculture Techniques; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology*; D005353:Fibronectins / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D008578:Meninges / Q000166:cytology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009433:Neural Inhibition / Q000187:drug effects / Q000502:physiology*; D016501:Neurites / Q000187:drug effects / Q000502:physiology; D009474:Neurons / Q000166:cytology*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism; D011720:Pyrazoles / Q000494:pharmacology; D011758:Pyrroles / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D036183:Receptor, EphB2 / Q000378:metabolism; D040121:Semaphorin-3A / Q000378:metabolism; D019063:Tenascin / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000037:antagonists & inhibitors / Q000494:pharmacology; D000099216:Chondroitin Sulfate Proteoglycan 4; D011804:Quinolines
D000941:Antigens; D001973:Bromodeoxyuridine; D002809:Chondroitin Sulfates; D005353:Fibronectins; D009419:Nerve Tissue Proteins; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D011509:Proteoglycans; D011720:Pyrazoles; D011758:Pyrroles; D036183:Receptor, EphB2; D040121:Semaphorin-3A; D019063:Tenascin; D053773:Transforming Growth Factor beta1; D000099216:Chondroitin Sulfate Proteoglycan 4; C506615:HTS 466284; D011804:Quinolines; C488218:Sema3a protein, rat