Development of a Functional Schwann Cell Phenotype from Autologous Porcine Bone Marrow Mononuclear Cells for Nerve Repair
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 sim...
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| Format: | Article |
| Language: | English |
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Wiley
2012-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2012/738484 |
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| author | Michael J. Rutten Michael Ann Janes Ivy R. Chang Cynthia R. Gregory Kenton W. Gregory |
| author_facet | Michael J. Rutten Michael Ann Janes Ivy R. Chang Cynthia R. Gregory Kenton W. Gregory |
| author_sort | Michael J. Rutten |
| collection | DOAJ |
| description | 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, 4 mM 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 (1–25 nM) increased p75(NGF) levels at 24–48 hrs. We found ATP dose-dependently increased intracellular calcium [Ca2+]i, with nucleotide potency being UTP=ATP>ADP>AMP>adenosine. Suramin blocked the ATP-induced [Ca2+]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 [Ca2+]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. |
| format | Article |
| id | doaj-art-47b10c5fd68b4635a6fd2d5b4a483e60 |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Wiley |
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| series | Stem Cells International |
| spelling | doaj-art-47b10c5fd68b4635a6fd2d5b4a483e602025-02-03T01:02:19ZengWileyStem Cells International1687-966X1687-96782012-01-01201210.1155/2012/738484738484Development of a Functional Schwann Cell Phenotype from Autologous Porcine Bone Marrow Mononuclear Cells for Nerve RepairMichael J. Rutten0Michael Ann Janes1Ivy R. Chang2Cynthia R. Gregory3Kenton W. Gregory4Providence Health and Services, 9555 SW Barnes Rd., Portland, OR 97225, USAProvidence Health and Services, 9555 SW Barnes Rd., Portland, OR 97225, USAProvidence Health and Services, 9555 SW Barnes Rd., Portland, OR 97225, USAOHSU Center for Regenerative Medicine, Oregon Health & Science University, 3181 S.W. Sam Jackson Park Road, Portland, OR 97239, USAProvidence Health and Services, 9555 SW Barnes Rd., Portland, OR 97225, USAAdult 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, 4 mM 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 (1–25 nM) increased p75(NGF) levels at 24–48 hrs. We found ATP dose-dependently increased intracellular calcium [Ca2+]i, with nucleotide potency being UTP=ATP>ADP>AMP>adenosine. Suramin blocked the ATP-induced [Ca2+]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 [Ca2+]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.http://dx.doi.org/10.1155/2012/738484 |
| spellingShingle | Michael J. Rutten Michael Ann Janes Ivy R. Chang Cynthia R. Gregory Kenton W. Gregory Development of a Functional Schwann Cell Phenotype from Autologous Porcine Bone Marrow Mononuclear Cells for Nerve Repair Stem Cells International |
| title | Development of a Functional Schwann Cell Phenotype from Autologous Porcine Bone Marrow Mononuclear Cells for Nerve Repair |
| title_full | Development of a Functional Schwann Cell Phenotype from Autologous Porcine Bone Marrow Mononuclear Cells for Nerve Repair |
| title_fullStr | Development of a Functional Schwann Cell Phenotype from Autologous Porcine Bone Marrow Mononuclear Cells for Nerve Repair |
| title_full_unstemmed | Development of a Functional Schwann Cell Phenotype from Autologous Porcine Bone Marrow Mononuclear Cells for Nerve Repair |
| title_short | Development of a Functional Schwann Cell Phenotype from Autologous Porcine Bone Marrow Mononuclear Cells for Nerve Repair |
| title_sort | development of a functional schwann cell phenotype from autologous porcine bone marrow mononuclear cells for nerve repair |
| url | http://dx.doi.org/10.1155/2012/738484 |
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