Live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride
The final stage of in vitro osteogenic differentiation is characterized by the production of mineral deposits containing calcium cations and inorganic phosphates, which populate the extracellular matrix surrounding the cell monolayer. Conventional histological techniques for the assessment of minera...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Mary Ann Liebert
2018
|
| Subjects: | |
| Online Access: | https://eprints.nottingham.ac.uk/49760/ |
| _version_ | 1848798071916331008 |
|---|---|
| author | Macri-Pellizzeri, Laura De Melo, Nigel Ahmed, Ifty Grant, David M. Scammell, Brigitte Sottile, Virginie |
| author_facet | Macri-Pellizzeri, Laura De Melo, Nigel Ahmed, Ifty Grant, David M. Scammell, Brigitte Sottile, Virginie |
| author_sort | Macri-Pellizzeri, Laura |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | The final stage of in vitro osteogenic differentiation is characterized by the production of mineral deposits containing calcium cations and inorganic phosphates, which populate the extracellular matrix surrounding the cell monolayer. Conventional histological techniques for the assessment of mineralization, such as Von Kossa and Alizarin Red S staining, are end-point techniques requiring cell fixation. Moreover, in both cases staining quantitation requires dye extraction which irreversibly alters the ECM conformation and structure, therefore preventing the use of the sample for further analysis.
In this study, the use of Tetracycline hydrochloride (TC) is proposed for the non-destructive staining, quantitation and imaging of mineralizing bone-like nodules in live cultures of human bone marrow mesenchymal stem cells (MSCs) cultured under osteogenic conditions. Overnight administration of TC to living cells was shown not to alter the metabolic activity or the progression of cell differentiation. When applied to differentiating cultures, cell exposure to serial doses of TC was found to produce quantifiable fluorescence emission specifically in osteogenic cultures. Incubation with TC enabled fluorescence imaging of mineralised areas in live cultures and the combination with other fluorophores using appropriate filters. These results demonstrate that serial TC administration over the differentiation time course provides a qualitative and quantitative tool for the monitoring and evaluation of the differentiation process in live cells. |
| first_indexed | 2025-11-14T20:13:57Z |
| format | Article |
| id | nottingham-49760 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T20:13:57Z |
| publishDate | 2018 |
| publisher | Mary Ann Liebert |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-497602020-05-08T09:30:14Z https://eprints.nottingham.ac.uk/49760/ Live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride Macri-Pellizzeri, Laura De Melo, Nigel Ahmed, Ifty Grant, David M. Scammell, Brigitte Sottile, Virginie The final stage of in vitro osteogenic differentiation is characterized by the production of mineral deposits containing calcium cations and inorganic phosphates, which populate the extracellular matrix surrounding the cell monolayer. Conventional histological techniques for the assessment of mineralization, such as Von Kossa and Alizarin Red S staining, are end-point techniques requiring cell fixation. Moreover, in both cases staining quantitation requires dye extraction which irreversibly alters the ECM conformation and structure, therefore preventing the use of the sample for further analysis. In this study, the use of Tetracycline hydrochloride (TC) is proposed for the non-destructive staining, quantitation and imaging of mineralizing bone-like nodules in live cultures of human bone marrow mesenchymal stem cells (MSCs) cultured under osteogenic conditions. Overnight administration of TC to living cells was shown not to alter the metabolic activity or the progression of cell differentiation. When applied to differentiating cultures, cell exposure to serial doses of TC was found to produce quantifiable fluorescence emission specifically in osteogenic cultures. Incubation with TC enabled fluorescence imaging of mineralised areas in live cultures and the combination with other fluorophores using appropriate filters. These results demonstrate that serial TC administration over the differentiation time course provides a qualitative and quantitative tool for the monitoring and evaluation of the differentiation process in live cells. Mary Ann Liebert 2018-03-30 Article PeerReviewed application/pdf en cc_by https://eprints.nottingham.ac.uk/49760/8/ten.tec.2017.0400.pdf Macri-Pellizzeri, Laura, De Melo, Nigel, Ahmed, Ifty, Grant, David M., Scammell, Brigitte and Sottile, Virginie (2018) Live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride. Tissue Engineering Part C: Methods, 24 (3). pp. 171-178. ISSN 1937-3384 stem cells osteogenesis in vitro differentiation live cell imaging quantitative assay https://www.liebertpub.com/doi/10.1089/ten.tec.2017.0400 doi:10.1089/ten.TEC.2017.0400 doi:10.1089/ten.TEC.2017.0400 |
| spellingShingle | stem cells osteogenesis in vitro differentiation live cell imaging quantitative assay Macri-Pellizzeri, Laura De Melo, Nigel Ahmed, Ifty Grant, David M. Scammell, Brigitte Sottile, Virginie Live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride |
| title | Live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride |
| title_full | Live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride |
| title_fullStr | Live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride |
| title_full_unstemmed | Live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride |
| title_short | Live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride |
| title_sort | live quantitative monitoring of mineral deposition in stem cells using tetracycline hydrochloride |
| topic | stem cells osteogenesis in vitro differentiation live cell imaging quantitative assay |
| url | https://eprints.nottingham.ac.uk/49760/ https://eprints.nottingham.ac.uk/49760/ https://eprints.nottingham.ac.uk/49760/ |