Gene-Modified Mesenchymal Stem Cells Express Functionally Active Nerve Growth Factor on an Engineered Poly Lactic Glycolic Acid (PLGA) Substrate
Delivery of cellular and/or trophic factors to the site of injury may promote neural repair or regeneration and return of function after peripheral nerve or spinal cord injury. Engineered scaffolds provide a platform to deliver therapeutic cells and neurotrophic molecules. We have genetically engine...
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| Veröffentlicht in: | Tissue engineering. Part A 2008-05, Vol.14 (5), p.681-690 |
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| container_title | Tissue engineering. Part A |
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| creator | Rooney, Gemma E. Moran, Cathal McMahon, Siobhan S. Ritter, Thomas Maenz, Martin Flügel, Alexander Dockery, Peter O'Brien, Timothy Howard, Linda Windebank, Anthony J. Barry, Frank P. |
| description | Delivery of cellular and/or trophic factors to the site of injury may promote neural repair or regeneration and return of function after peripheral nerve or spinal cord injury. Engineered scaffolds provide a platform to deliver therapeutic cells and neurotrophic molecules. We have genetically engineered mesenchymal stem cells (MSCs) from the green rat (CZ-004 [SD TgN(act-EGFP)OsbCZ-004]) to express nerve growth factor (NGF) using an adenoviral vector. Cells maintained their stem cell phenotype as judged by expression of CD71 and CD172 markers, and absence of the hematopoietic marker CD45. Cells continued to express green fluorescent protein (GFP) on a long-term basis. Morphology, viability, and growth kinetics were maintained when cells were grown on a poly-lactic-co-glycolic acid (PLGA) polymer scaffold. Under appropriate growth conditions, they differentiated into chondrogenic, osteogenic, and adipogenic phenotypes, demonstrating that they retained their characteristics as MSCs. NGF was secreted from transduced MSCs at physiologically relevant levels (∼25 ng/mL) measured by enzyme-linked immunoabsorbent assay (ELISA). Secreted NGF was functionally active in a neurite growth assay with PC12 cells. We conclude that MSCs are a good candidate for delivery of therapeutic factors into the injured nervous system. They are autologous, may be genetically modified to express neurotrophins, and are compatible with polymer surfaces that may be used as a potential delivery system. |
| doi_str_mv | 10.1089/tea.2007.0260 |
| format | Article |
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Engineered scaffolds provide a platform to deliver therapeutic cells and neurotrophic molecules. We have genetically engineered mesenchymal stem cells (MSCs) from the green rat (CZ-004 [SD TgN(act-EGFP)OsbCZ-004]) to express nerve growth factor (NGF) using an adenoviral vector. Cells maintained their stem cell phenotype as judged by expression of CD71 and CD172 markers, and absence of the hematopoietic marker CD45. Cells continued to express green fluorescent protein (GFP) on a long-term basis. Morphology, viability, and growth kinetics were maintained when cells were grown on a poly-lactic-co-glycolic acid (PLGA) polymer scaffold. Under appropriate growth conditions, they differentiated into chondrogenic, osteogenic, and adipogenic phenotypes, demonstrating that they retained their characteristics as MSCs. NGF was secreted from transduced MSCs at physiologically relevant levels (∼25 ng/mL) measured by enzyme-linked immunoabsorbent assay (ELISA). Secreted NGF was functionally active in a neurite growth assay with PC12 cells. We conclude that MSCs are a good candidate for delivery of therapeutic factors into the injured nervous system. They are autologous, may be genetically modified to express neurotrophins, and are compatible with polymer surfaces that may be used as a potential delivery system.</description><identifier>ISSN: 1937-3341</identifier><identifier>EISSN: 1937-335X</identifier><identifier>DOI: 10.1089/tea.2007.0260</identifier><identifier>PMID: 18402551</identifier><language>eng</language><publisher>United States: Mary Ann Liebert, Inc</publisher><subject>Animals ; Animals, Genetically Modified ; Biocompatible Materials ; Biomedical materials ; Care and treatment ; Cellular biology ; Female ; Gene expression ; Genetic aspects ; Green Fluorescent Proteins - genetics ; Green Fluorescent Proteins - metabolism ; Health aspects ; Lactic Acid ; Male ; Materials Testing ; Measurement ; Mesenchymal Stromal Cells - metabolism ; Nerve growth factor ; Nerve Growth Factor - genetics ; Nerve Growth Factor - metabolism ; Nervous system diseases ; Neurites - ultrastructure ; Neurons ; Physiological aspects ; Polyglycolic Acid ; Proteins ; Rats ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; Stem cells ; Tissue engineering ; Tissue Engineering - methods ; Tissue Scaffolds ; Transduction, Genetic</subject><ispartof>Tissue engineering. 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Part A</title><addtitle>Tissue Eng Part A</addtitle><description>Delivery of cellular and/or trophic factors to the site of injury may promote neural repair or regeneration and return of function after peripheral nerve or spinal cord injury. Engineered scaffolds provide a platform to deliver therapeutic cells and neurotrophic molecules. We have genetically engineered mesenchymal stem cells (MSCs) from the green rat (CZ-004 [SD TgN(act-EGFP)OsbCZ-004]) to express nerve growth factor (NGF) using an adenoviral vector. Cells maintained their stem cell phenotype as judged by expression of CD71 and CD172 markers, and absence of the hematopoietic marker CD45. Cells continued to express green fluorescent protein (GFP) on a long-term basis. Morphology, viability, and growth kinetics were maintained when cells were grown on a poly-lactic-co-glycolic acid (PLGA) polymer scaffold. Under appropriate growth conditions, they differentiated into chondrogenic, osteogenic, and adipogenic phenotypes, demonstrating that they retained their characteristics as MSCs. NGF was secreted from transduced MSCs at physiologically relevant levels (∼25 ng/mL) measured by enzyme-linked immunoabsorbent assay (ELISA). Secreted NGF was functionally active in a neurite growth assay with PC12 cells. We conclude that MSCs are a good candidate for delivery of therapeutic factors into the injured nervous system. They are autologous, may be genetically modified to express neurotrophins, and are compatible with polymer surfaces that may be used as a potential delivery system.</description><subject>Animals</subject><subject>Animals, Genetically Modified</subject><subject>Biocompatible Materials</subject><subject>Biomedical materials</subject><subject>Care and treatment</subject><subject>Cellular biology</subject><subject>Female</subject><subject>Gene expression</subject><subject>Genetic aspects</subject><subject>Green Fluorescent Proteins - genetics</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>Health aspects</subject><subject>Lactic Acid</subject><subject>Male</subject><subject>Materials Testing</subject><subject>Measurement</subject><subject>Mesenchymal Stromal Cells - metabolism</subject><subject>Nerve growth factor</subject><subject>Nerve Growth Factor - genetics</subject><subject>Nerve Growth Factor - metabolism</subject><subject>Nervous system diseases</subject><subject>Neurites - ultrastructure</subject><subject>Neurons</subject><subject>Physiological aspects</subject><subject>Polyglycolic Acid</subject><subject>Proteins</subject><subject>Rats</subject><subject>Recombinant Proteins - genetics</subject><subject>Recombinant Proteins - metabolism</subject><subject>Stem cells</subject><subject>Tissue engineering</subject><subject>Tissue Engineering - methods</subject><subject>Tissue Scaffolds</subject><subject>Transduction, Genetic</subject><issn>1937-3341</issn><issn>1937-335X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqFkl2L1DAUhoso7odeeitBQfSiY5K2SXtZhpkqzOrCKngXMunJbpY0mU1a3fkX_mRTZlAUYQnkHE6e8yY5vFn2guAFwXXzfgS5oBjzBaYMP8pOSVPwvCiqb49_5yU5yc5ivMWYYcb50-yE1CWmVUVOs58dOMgvfG-0gR5dQASnbvaDtOhqhAEtwdqIVve7ADGi9eTUaLyT1u5Rm9LvgD5BSHsX_I_xBq2lGn1A3iHp0MpdGwcQku6lTw2bdGgU6uxeeZuSVpkevb3cdO07dDVt4xjkCM-yJ1raCM-P8Tz7ul59WX7IN5-7j8t2k6uKsjHXhEjOel41DdO6ZE3PGZljXcqSKF7VBaskloTzhvZlj7dUypLqYqtxRVVdnGdvDrq74O8miKMYTFTpt9KBn6JgTUVKgvmDYBoyKQhvEvjqH_DWTyHNKgqKCae8Lme11wfoWloQxmmffq1mRdEmkXQra0iiFv-h0uphMMo70CbV_2rIDw0q-BgDaLELZpBhLwgWs09E8omYfSJmnyT-5fGt03aA_g99NEYCigMwl6Vz1sAWwviA7C-om8dS</recordid><startdate>20080501</startdate><enddate>20080501</enddate><creator>Rooney, Gemma E.</creator><creator>Moran, Cathal</creator><creator>McMahon, Siobhan S.</creator><creator>Ritter, Thomas</creator><creator>Maenz, Martin</creator><creator>Flügel, Alexander</creator><creator>Dockery, Peter</creator><creator>O'Brien, Timothy</creator><creator>Howard, Linda</creator><creator>Windebank, Anthony J.</creator><creator>Barry, Frank P.</creator><general>Mary Ann Liebert, Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QP</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>88I</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>M7P</scope><scope>PHGZM</scope><scope>PHGZT</scope><scope>PJZUB</scope><scope>PKEHL</scope><scope>PPXIY</scope><scope>PQEST</scope><scope>PQGLB</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20080501</creationdate><title>Gene-Modified Mesenchymal Stem Cells Express Functionally Active Nerve Growth Factor on an Engineered Poly Lactic Glycolic Acid (PLGA) Substrate</title><author>Rooney, Gemma E. ; Moran, Cathal ; McMahon, Siobhan S. ; Ritter, Thomas ; Maenz, Martin ; Flügel, Alexander ; Dockery, Peter ; O'Brien, Timothy ; Howard, Linda ; Windebank, Anthony J. ; Barry, Frank P.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c526t-f11a76d75996ff469d761f46984a41c758365a0a17792d4d0b2aa42f3bf052c83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Animals</topic><topic>Animals, Genetically Modified</topic><topic>Biocompatible Materials</topic><topic>Biomedical materials</topic><topic>Care and treatment</topic><topic>Cellular biology</topic><topic>Female</topic><topic>Gene expression</topic><topic>Genetic aspects</topic><topic>Green Fluorescent Proteins - 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Part A</jtitle><addtitle>Tissue Eng Part A</addtitle><date>2008-05-01</date><risdate>2008</risdate><volume>14</volume><issue>5</issue><spage>681</spage><epage>690</epage><pages>681-690</pages><issn>1937-3341</issn><eissn>1937-335X</eissn><abstract>Delivery of cellular and/or trophic factors to the site of injury may promote neural repair or regeneration and return of function after peripheral nerve or spinal cord injury. Engineered scaffolds provide a platform to deliver therapeutic cells and neurotrophic molecules. We have genetically engineered mesenchymal stem cells (MSCs) from the green rat (CZ-004 [SD TgN(act-EGFP)OsbCZ-004]) to express nerve growth factor (NGF) using an adenoviral vector. Cells maintained their stem cell phenotype as judged by expression of CD71 and CD172 markers, and absence of the hematopoietic marker CD45. Cells continued to express green fluorescent protein (GFP) on a long-term basis. Morphology, viability, and growth kinetics were maintained when cells were grown on a poly-lactic-co-glycolic acid (PLGA) polymer scaffold. Under appropriate growth conditions, they differentiated into chondrogenic, osteogenic, and adipogenic phenotypes, demonstrating that they retained their characteristics as MSCs. NGF was secreted from transduced MSCs at physiologically relevant levels (∼25 ng/mL) measured by enzyme-linked immunoabsorbent assay (ELISA). Secreted NGF was functionally active in a neurite growth assay with PC12 cells. We conclude that MSCs are a good candidate for delivery of therapeutic factors into the injured nervous system. They are autologous, may be genetically modified to express neurotrophins, and are compatible with polymer surfaces that may be used as a potential delivery system.</abstract><cop>United States</cop><pub>Mary Ann Liebert, Inc</pub><pmid>18402551</pmid><doi>10.1089/tea.2007.0260</doi><tpages>10</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1937-3341 |
| ispartof | Tissue engineering. Part A, 2008-05, Vol.14 (5), p.681-690 |
| issn | 1937-3341 1937-335X |
| language | eng |
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| source | Mary Ann Liebert Online Subscription; MEDLINE; Alma/SFX Local Collection |
| subjects | Animals Animals, Genetically Modified Biocompatible Materials Biomedical materials Care and treatment Cellular biology Female Gene expression Genetic aspects Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism Health aspects Lactic Acid Male Materials Testing Measurement Mesenchymal Stromal Cells - metabolism Nerve growth factor Nerve Growth Factor - genetics Nerve Growth Factor - metabolism Nervous system diseases Neurites - ultrastructure Neurons Physiological aspects Polyglycolic Acid Proteins Rats Recombinant Proteins - genetics Recombinant Proteins - metabolism Stem cells Tissue engineering Tissue Engineering - methods Tissue Scaffolds Transduction, Genetic |
| title | Gene-Modified Mesenchymal Stem Cells Express Functionally Active Nerve Growth Factor on an Engineered Poly Lactic Glycolic Acid (PLGA) Substrate |
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