Development of a functional Schwann cell phenotype from autologous porcine bone marrow mononuclear cells for nerve repair

Michael J. Rutten, Michael Ann Janes, Ivy R. Chang, Cynthia R. Gregory, Kenton W. Gregory

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Adult bone marrow mononuclear cells (BM-MNCs) are a potential resource for making Schwann cells to repair damaged peripheral nerves. However, many methods of producing Schwann-like cells can be laborious with the cells lacking a functional phenotype. The objective of this study was to develop a simple and rapid method using autologous BM-MNCs to produce a phenotypic and functional Schwann-like cell. Adult porcine bone marrow was collected and enriched for BM-MNCs using a SEPAX device, then cells cultured in Neurobasal media, 4mM L-glutamine and 20% serum. After 6-8 days, the cultures expressed Schwann cell markers, S-100, O4, GFAP, were FluoroMyelin positive, but had low p75(NGF) expression. Addition of neuregulin (1-25nM) increased p75(NGF) levels at 24-48hrs. We found ATP dose-dependently increased intracellular calcium [Ca 2+] i, with nucleotide potency being UTP = ATP > ADP > AMP adenosine. Suramin blocked the ATP-induced [Ca 2+] i but α, β,-methylene-ATP had little effect suggesting an ATP purinergic P2Y2 G-protein-coupled receptor is present. Both the Schwann cell markers and ATP-induced [Ca 2+] i sensitivity decreased in cells passaged >20 times. Our studies indicate that autologous BM-MNCs can be induced to form a phenotypic and functional Schwann-like cell which could be used for peripheral nerve repair.

Original languageEnglish (US)
Article number738484
JournalStem Cells International
DOIs
StatePublished - 2012

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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