99mTc-methyl-diphosphonate binding to mineral deposits in cultures of marrow-derived mesenchymal stem cells in osteogenic medium
Osteogenic differentiation of adult mesenchymal stem cells (MSCs) is a promising method for the therapy of critical size bone defects and other applications, drawing attention to the importance of in vitro assays for its evaluation. While there are standardized protocols to induce osteogenic differe...
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| Hauptverfasser: | , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
5 Apr 2019
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| In: |
Tissue engineering. Part C, Methods
Year: 2019, Jahrgang: 25, Heft: 1, Pages: 49-57 |
| ISSN: | 1937-3392 |
| DOI: | 10.1089/ten.tec.2018.0299 |
| Online-Zugang: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1089/ten.tec.2018.0299 Verlag, lizenzpflichtig, Volltext: https://www.liebertpub.com/doi/10.1089/ten.tec.2018.0299 
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| Verfasserangaben: | Tobias Grossner, Tobias Gotterbarm, Victor H. Gerbaudo, Uwe Haberkorn, Myron Spector |
MARC
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| 520 | |a Osteogenic differentiation of adult mesenchymal stem cells (MSCs) is a promising method for the therapy of critical size bone defects and other applications, drawing attention to the importance of in vitro assays for its evaluation. While there are standardized protocols to induce osteogenic differentiation in monolayer cultures, there is a lack of validated nondestructive procedures to quantify the osteogenic potential of MSCs directly by accessing the mineral deposition. In this study, we present a new method to determine the osteogenic potential of MSCs using the radioactive tracer, 99mTechnetium-methylene diphosphonate (99mTc-MDP), to bind in vitro to newly produced mineral deposits from osteogenic-induced stem cells. Thirty five-millimeter Petri dishes were seeded with goat and human MSCs and differentiated into the osteogenic lineage. 99mTc-MDP was applied to each dish, and acquisition of the bound tracer was imaged with a gamma camera. The results revealed a significantly higher uptake in the osteogenic-differentiated MSCs for both goat and human cells (p < 0.008 goat; 0.01 human), compared with controls grown in expansion medium. We were able to show that 99mTc-MDP labeling is a suitable method to quantify the amount of extracellular mineral deposited by MSCs. Goat and human MSCs displayed similar uptake characteristics (Spearman-Rho Correlation coefficient r = 0.603). The radionuclide method was validated by quantitative Alizarin Red staining, which showed a high correlation (Spearman-Rho Correlation coefficient r = 0.668). This radionuclide method opens a new approach for evaluating osteogenic potential of MSCs and for longitudinal studies of the process. - - Impact Statement - - The work is notable for describing a highly sensitive, quantitative, and nondestructive method for evaluating the in vitro amount of mineral accompanying different types of osteogenic differentiation of mesenchymal stem cells in a monolayer cell culture. What is so unique and useful about the method is that it has the potential to be used to define the kinetics of the differentiation process, reflected in the mineralization, without destroying the monolayer. Therefore, it remains intact for further experiments. | ||
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