Transient 100 nM dexamethasone treatment reduces inter- and intraindividual variations in osteoblastic differentiation of bone marrow-derived human mesenchymal stem cells
| Autor: | H. Kalervo Väänänen, Terhi J. Heino, Jessica J. Alm, Teuvo Hentunen, Hannu T. Aro |
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| Rok vydání: | 2012 |
| Předmět: |
Adult
medicine.medical_specialty Cell Survival Cellular differentiation Osteocalcin Biomedical Engineering Cell Culture Techniques Medicine (miscellaneous) Bioengineering Apoptosis Bone Marrow Cells Collagen Type I Dexamethasone Internal medicine polycyclic compounds medicine Humans Viability assay Glucocorticoids Cell Proliferation ta3126 Osteoblasts biology Tissue Engineering Cell growth Chemistry Mesenchymal stem cell Cell Differentiation Mesenchymal Stem Cells Middle Aged Alkaline Phosphatase Endocrinology medicine.anatomical_structure biology.protein Alkaline phosphatase Female Bone marrow hormones hormone substitutes and hormone antagonists |
| Zdroj: | Tissue Engineering Part C Methods. 18(9):658-666 |
| ISSN: | 1937-3384 |
| DOI: | 10.1089/ten.TEC.2011.0675 |
| Popis: | The development of in vitro culturing techniques for osteoblastic differentiation of human mesenchymal stem cells (hMSC) is important for cell biology research and the development of tissue-engineering applications. Dexamethasone (Dex) is a commonly used supplement, but the optimal use of Dex treatment is still unclear. By adjusting the timing of Dex supplementation, the negative effects of long-term Dex treatment could be overcome. Transient Dex treatment could contribute toward minimizing broad donor variation, which is a major challenge. We compared the two most widely used Dex concentrations of 10 and 100 nM as transient or continuous treatment and studied inter- and intraindividual variations in osteoblastic differentiation of hMSC. Characterized bone marrow-derived hMSC from 17 female donors of different age groups were used. During osteoblastic induction, the cells were treated with 10 or 100 nM Dex either transiently for different time periods or continuously. Differentiation was evaluated by measuring alkaline phosphatase (ALP) activity and staining for ALP, von Kossa, collagen type I, and osteocalcin. Cell proliferation, cell viability, and apoptosis were also monitored. The strongest osteoblastic differentiation was observed when 100 nM Dex was present for the first week. In terms of inter- and intraindividual coefficients of variations, transient treatment with 100 nM Dex was superior to the other culture conditions and showed the lowest variations in all age groups. This study demonstrates that the temporary presence of 100 nM Dex during the first week of induction culture promotes hMSC osteoblastic differentiation and reduces inter- and intraindividual variations. With this protocol, we can reproducibly produce functional osteoblasts in vitro from the hMSC of different donor populations. |
| Databáze: | OpenAIRE |
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