Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System
Intervertebral disc (IVD) cell therapy with unconditioned 2D expanded mesenchymal stem cells (MSC) is a promising concept yet challenging to realize. Differentiation of MSCs by nonviral gene delivery of growth and differentiation factor 5 (GDF5) by electroporation mediated gene transfer could be an...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2013-01-01
|
| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2013/326828 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832559783541997568 |
|---|---|
| author | Christian Bucher Amiq Gazdhar Lorin M. Benneker Thomas Geiser Benjamin Gantenbein-Ritter |
| author_facet | Christian Bucher Amiq Gazdhar Lorin M. Benneker Thomas Geiser Benjamin Gantenbein-Ritter |
| author_sort | Christian Bucher |
| collection | DOAJ |
| description | Intervertebral disc (IVD) cell therapy with unconditioned 2D expanded mesenchymal stem cells (MSC) is a promising concept yet challenging to realize. Differentiation of MSCs by nonviral gene delivery of growth and differentiation factor 5 (GDF5) by electroporation mediated gene transfer could be an excellent source for cell transplantation. Human MSCs were harvested from bone marrow aspirate and GDF5 gene transfer was achieved by in vitro electroporation. Transfected cells were cultured as monolayers and as 3D cultures in 1.2% alginate bead culture. MSC expressed GDF5 efficiently for up to 21 days. The combination of GDF5 gene transfer and 3D culture in alginate showed an upregulation of aggrecan and SOX9, two markers for chondrogenesis, and KRT19 as a marker for discogenesis compared to untransfected cells. The cells encapsulated in alginate produced more proteoglycans expressed in GAG/DNA ratio. Furthermore, GDF5 transfected MCS injected into an IVD papain degeneration organ culture model showed a partial recovery of the GAG/DNA ratio after 7 days. In this study we demonstrate the potential of GDF5 transfected MSC as a promising approach for clinical translation for disc regeneration. |
| format | Article |
| id | doaj-art-a9d864be3cdb47eba8a5be65de77f5f4 |
| institution | Kabale University |
| issn | 1687-966X 1687-9678 |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Stem Cells International |
| spelling | doaj-art-a9d864be3cdb47eba8a5be65de77f5f42025-02-03T01:29:14ZengWileyStem Cells International1687-966X1687-96782013-01-01201310.1155/2013/326828326828Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture SystemChristian Bucher0Amiq Gazdhar1Lorin M. Benneker2Thomas Geiser3Benjamin Gantenbein-Ritter4Institute for Surgical Technology and Biomechanics, Tissue & Organ Mechanobiology, Medical Faculty, University of Bern, Stauffacherstraße 78, 3014 Bern, SwitzerlandDepartment of Pulmonary Medicine, University Hospital Bern and Department for Clinical Research, University of Bern, 3010 Bern, SwitzerlandDepartment of Orthopaedic Surgery, Insel Hospital, University of Bern, 3014 Bern, SwitzerlandDepartment of Pulmonary Medicine, University Hospital Bern and Department for Clinical Research, University of Bern, 3010 Bern, SwitzerlandInstitute for Surgical Technology and Biomechanics, Tissue & Organ Mechanobiology, Medical Faculty, University of Bern, Stauffacherstraße 78, 3014 Bern, SwitzerlandIntervertebral disc (IVD) cell therapy with unconditioned 2D expanded mesenchymal stem cells (MSC) is a promising concept yet challenging to realize. Differentiation of MSCs by nonviral gene delivery of growth and differentiation factor 5 (GDF5) by electroporation mediated gene transfer could be an excellent source for cell transplantation. Human MSCs were harvested from bone marrow aspirate and GDF5 gene transfer was achieved by in vitro electroporation. Transfected cells were cultured as monolayers and as 3D cultures in 1.2% alginate bead culture. MSC expressed GDF5 efficiently for up to 21 days. The combination of GDF5 gene transfer and 3D culture in alginate showed an upregulation of aggrecan and SOX9, two markers for chondrogenesis, and KRT19 as a marker for discogenesis compared to untransfected cells. The cells encapsulated in alginate produced more proteoglycans expressed in GAG/DNA ratio. Furthermore, GDF5 transfected MCS injected into an IVD papain degeneration organ culture model showed a partial recovery of the GAG/DNA ratio after 7 days. In this study we demonstrate the potential of GDF5 transfected MSC as a promising approach for clinical translation for disc regeneration.http://dx.doi.org/10.1155/2013/326828 |
| spellingShingle | Christian Bucher Amiq Gazdhar Lorin M. Benneker Thomas Geiser Benjamin Gantenbein-Ritter Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System Stem Cells International |
| title | Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System |
| title_full | Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System |
| title_fullStr | Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System |
| title_full_unstemmed | Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System |
| title_short | Nonviral Gene Delivery of Growth and Differentiation Factor 5 to Human Mesenchymal Stem Cells Injected into a 3D Bovine Intervertebral Disc Organ Culture System |
| title_sort | nonviral gene delivery of growth and differentiation factor 5 to human mesenchymal stem cells injected into a 3d bovine intervertebral disc organ culture system |
| url | http://dx.doi.org/10.1155/2013/326828 |
| work_keys_str_mv | AT christianbucher nonviralgenedeliveryofgrowthanddifferentiationfactor5tohumanmesenchymalstemcellsinjectedintoa3dbovineintervertebraldiscorganculturesystem AT amiqgazdhar nonviralgenedeliveryofgrowthanddifferentiationfactor5tohumanmesenchymalstemcellsinjectedintoa3dbovineintervertebraldiscorganculturesystem AT lorinmbenneker nonviralgenedeliveryofgrowthanddifferentiationfactor5tohumanmesenchymalstemcellsinjectedintoa3dbovineintervertebraldiscorganculturesystem AT thomasgeiser nonviralgenedeliveryofgrowthanddifferentiationfactor5tohumanmesenchymalstemcellsinjectedintoa3dbovineintervertebraldiscorganculturesystem AT benjamingantenbeinritter nonviralgenedeliveryofgrowthanddifferentiationfactor5tohumanmesenchymalstemcellsinjectedintoa3dbovineintervertebraldiscorganculturesystem |