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We have used PC12 cells to examine the effects of factors secreted by Schwann cells that promote cell survival and neurite outgrowth, and hence are likely candidates for promoting neuronal regeneration. RT-PCR showed that primary Schwann cells produced a range of neurotrophins, excluding NT3, but this profile was different from either of two cell lines SCTM41 or PVGSCSV40T, or forskolin-expanded Schwann cells. The effects of Schwann cell conditioned media on neurite outgrowth was tested against a range of factors, and showed clear neuritogenic effects. Of the factors tested, only NGF had a significant response on neuritogenesis. Western blotting for neurofilaments showed that primary Schwann cells induced a strong response close to that of NGF. The Trk tyrosine kinase inhibitor K252a did not block the neuritogenic effects of primary Schwann cells. In contrast, K252a blocked both NGF and the SCTM41 cell effects. Schwann cell conditioned media also enhanced PC12 cell survival. Again, in contrast with NGF or SCTM41 cells, the primary Schwann cell effect was Trk tyrosine kinase independent. The Schwann cell conditioned medium contains a protein factor (greater than 12 kDa and broken down by trypsin treatment) with remarkable thermal stability (unaffected at 95 degrees C for 15 min) and the ability to bind heparin. Our results provide clear evidence that Schwann cells produce factors other than those already known to stimulate a neural phenotype in PC12 cells, and which thus have potential regeneration enhancing effects.

Original publication

DOI

10.1002/neu.20119

Type

Journal article

Journal

J Neurobiol

Publication Date

04/2005

Volume

63

Pages

29 - 48

Keywords

Animals, Animals, Newborn, Blotting, Western, Carbazoles, Cell Survival, Cells, Cultured, Culture Media, Conditioned, Drug Interactions, Enzyme Inhibitors, Fibroblasts, Immunohistochemistry, Indole Alkaloids, Indoles, Nerve Growth Factors, Neurites, Neurofilament Proteins, PC12 Cells, RNA, Messenger, Rats, Reverse Transcriptase Polymerase Chain Reaction, Schwann Cells