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

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

Research output: Contribution to journalArticle

5 Citations (Scopus)

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

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Schwann Cells
Bone Marrow Cells
Swine
Adenosine Triphosphate
Phenotype
Nerve Growth Factor
Peripheral Nerves
Neuregulin-1
Suramin
Uridine Triphosphate
Adenosine Monophosphate
G-Protein-Coupled Receptors
Glutamine
Adenosine
Adenosine Diphosphate
Cultured Cells
Nucleotides
Bone Marrow
Calcium
Equipment and Supplies

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

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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.",
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AU - Rutten, Michael

AU - Janes, Michael Ann

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AU - Gregory, Cynthia

AU - Gregory, Kenton

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