Assessment of a Nuclear Affinity Labeling Method for Tracking Implanted Mesenchymal Stem Cells
Therapeutic implantation of mesenchymal stem cells (MSCs) is entering the realm of clinical trials for several human diseases, and yet much remains uncertain regarding their dynamic distribution and cell fate after in vivo application. Discrepancies in the literature can be attributed in part to the...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2008-08-01
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| Series: | Cell Transplantation |
| Online Access: | https://doi.org/10.3727/096368908786576444 |
| _version_ | 1818068394214162432 |
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| author | Merced Leiker Gen Suzuki Vijay S. Iyer Canty John M Techung Lee |
| author_facet | Merced Leiker Gen Suzuki Vijay S. Iyer Canty John M Techung Lee |
| author_sort | Merced Leiker |
| collection | DOAJ |
| description | Therapeutic implantation of mesenchymal stem cells (MSCs) is entering the realm of clinical trials for several human diseases, and yet much remains uncertain regarding their dynamic distribution and cell fate after in vivo application. Discrepancies in the literature can be attributed in part to the use of different cell labeling/tracking methods and cell administration protocols. To identify a stem cell detection method suitable for myocardial implantation in a large animal model, we experimented on three different MSC labeling methods: adenovirus-mediated expression of enhanced green fluorescence protein (EGFP) and β-galactosidase (LacZ), and nuclear staining with DAPI. Intramuscular and intracoronary administrations of labeled porcine MSCs identified the nuclear affinity dye to be a reliable stem cell tracking marker. Stem cell identification is facilitated by an optimized live cell labeling condition generating bright blue fluorescence sharply confined to the nucleus. DAPI-labeled MSCs retained full viability, ceased proliferation, and exhibited an increased differentiation potential. The labeled MSCs remained fully active in expressing key growth factor and cytokine genes, and notably exhibited enhanced expression of the chemokine receptor CXCR4 and its ligand SDF1, indicating their competency in response to tissue injury. Histological analysis revealed that approximately half a million MSCs or ~2% of the administered MSCs remained localized in the normal pig heart 2 weeks after coronary infusion. That the vast majority of these identified MSCs were interstitial indicated the ability of MSCs to migrate across the coronary endothelium. No evidence was obtained indicating MSC differentiation to cardiomyocyte. |
| first_indexed | 2024-12-10T15:38:53Z |
| format | Article |
| id | doaj.art-5a3b4e271de44219832d0d1e930876a9 |
| institution | Directory Open Access Journal |
| issn | 0963-6897 1555-3892 |
| language | English |
| last_indexed | 2024-12-10T15:38:53Z |
| publishDate | 2008-08-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Cell Transplantation |
| spelling | doaj.art-5a3b4e271de44219832d0d1e930876a92022-12-22T01:43:10ZengSAGE PublishingCell Transplantation0963-68971555-38922008-08-011710.3727/096368908786576444Assessment of a Nuclear Affinity Labeling Method for Tracking Implanted Mesenchymal Stem CellsMerced Leiker0Gen Suzuki1Vijay S. Iyer2Canty John M3Techung Lee4Center for Research in Cardiovascular Medicine, University at Buffalo, Buffalo, NY, USACenter for Research in Cardiovascular Medicine, University at Buffalo, Buffalo, NY, USACenter for Research in Cardiovascular Medicine, University at Buffalo, Buffalo, NY, USACenter for Research in Cardiovascular Medicine, University at Buffalo, Buffalo, NY, USACenter for Research in Cardiovascular Medicine, University at Buffalo, Buffalo, NY, USATherapeutic implantation of mesenchymal stem cells (MSCs) is entering the realm of clinical trials for several human diseases, and yet much remains uncertain regarding their dynamic distribution and cell fate after in vivo application. Discrepancies in the literature can be attributed in part to the use of different cell labeling/tracking methods and cell administration protocols. To identify a stem cell detection method suitable for myocardial implantation in a large animal model, we experimented on three different MSC labeling methods: adenovirus-mediated expression of enhanced green fluorescence protein (EGFP) and β-galactosidase (LacZ), and nuclear staining with DAPI. Intramuscular and intracoronary administrations of labeled porcine MSCs identified the nuclear affinity dye to be a reliable stem cell tracking marker. Stem cell identification is facilitated by an optimized live cell labeling condition generating bright blue fluorescence sharply confined to the nucleus. DAPI-labeled MSCs retained full viability, ceased proliferation, and exhibited an increased differentiation potential. The labeled MSCs remained fully active in expressing key growth factor and cytokine genes, and notably exhibited enhanced expression of the chemokine receptor CXCR4 and its ligand SDF1, indicating their competency in response to tissue injury. Histological analysis revealed that approximately half a million MSCs or ~2% of the administered MSCs remained localized in the normal pig heart 2 weeks after coronary infusion. That the vast majority of these identified MSCs were interstitial indicated the ability of MSCs to migrate across the coronary endothelium. No evidence was obtained indicating MSC differentiation to cardiomyocyte.https://doi.org/10.3727/096368908786576444 |
| spellingShingle | Merced Leiker Gen Suzuki Vijay S. Iyer Canty John M Techung Lee Assessment of a Nuclear Affinity Labeling Method for Tracking Implanted Mesenchymal Stem Cells Cell Transplantation |
| title | Assessment of a Nuclear Affinity Labeling Method for Tracking Implanted Mesenchymal Stem Cells |
| title_full | Assessment of a Nuclear Affinity Labeling Method for Tracking Implanted Mesenchymal Stem Cells |
| title_fullStr | Assessment of a Nuclear Affinity Labeling Method for Tracking Implanted Mesenchymal Stem Cells |
| title_full_unstemmed | Assessment of a Nuclear Affinity Labeling Method for Tracking Implanted Mesenchymal Stem Cells |
| title_short | Assessment of a Nuclear Affinity Labeling Method for Tracking Implanted Mesenchymal Stem Cells |
| title_sort | assessment of a nuclear affinity labeling method for tracking implanted mesenchymal stem cells |
| url | https://doi.org/10.3727/096368908786576444 |
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