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Gene Transfer of the Runx2 Transcription Factor Enhances Osteogenic Activity of Bone Marrow Stromal Cells in Vitro and in Vivo
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Gene Transfer of the Runx2 Transcription Factor Enhances Osteogenic Activity of Bone Marrow Stromal Cells in Vitro and in Vivo
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Journal Article
Gene Transfer of the Runx2 Transcription Factor Enhances Osteogenic Activity of Bone Marrow Stromal Cells in Vitro and in Vivo
2005
Overview
Marrow stromal cells (MSCs) have the potential to differentiate into multiple mesenchymal cell types. To harness the power of MSCs for bone regeneration, methods must be developed to direct their differentiation selectively to the osteoblast lineage. The objective of this study was to examine the feasibility of using ex vivo Runx2 gene transfer to enhance the osteogenic activity of MSCs. Primary MSCs isolated from C57BL6 mice were transduced with adenoviral vectors encoding beta-galactosidase or Runx2. Cells transduced with Ad-Runx2 expressed Runx2 protein and underwent osteoblast differentiation as measured by increases in alkaline phosphatase activity and mineralization. Time-course studies revealed that Runx2 protein was highest 1 day after transduction and declined below the limits of detection by 15 days. Osteoblast marker mRNA expression paralleled Runx2 levels. In contrast, Runx2-dependent mineralization persisted for the duration of the experiment. To assess in vivo osteogenic activity, Ad-Runx2-transduced and control MSCs were adsorbed to two different carrier scaffolds and subcutaneously implanted into C57BL6 mice. In both cases, MSCs expressing Runx2 formed substantially more bone than cells transduced with control virus. Taken together, these studies indicate that Runx2 gene transfer may be an effective route to enhance the osteogenic potential of MSCs.
Publisher
Elsevier Limited
Subject
/ Animals
/ Bone Marrow Cells - drug effects
/ Bone Marrow Cells - metabolism
/ Bone Regeneration - drug effects
/ Core Binding Factor Alpha 1 Subunit - genetics
/ Core Binding Factor Alpha 1 Subunit - metabolism
/ Core Binding Factor Alpha 1 Subunit - pharmacology
/ Mice
/ Proteins
/ Reverse Transcriptase Polymerase Chain Reaction
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