Live Simultaneous Monitoring of Mineral Deposition and Lipid Accumulation in Differentiating Stem Cells
| Autor: | Michael E. Symonds, Laura Macri-Pellizzeri, Ifty Ahmed, Nigel De Melo, Virginie Sottile, Robert Markus, Sarah Mcginlay |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
mineralisation
Biomedical Engineering 030209 endocrinology & metabolism Bioengineering Biochemistry lcsh:Technology Article Biomaterials 03 medical and health sciences 0302 clinical medicine medicine Progenitor cell 030304 developmental biology 0303 health sciences Chemistry lcsh:T Mesenchymal stem cell Osteoblast Cell biology Staining stem cell medicine.anatomical_structure Vital stain Adipogenesis Molecular Medicine Bone marrow Stem cell live monitoring lipid detection Biotechnology |
| Zdroj: | Biomimetics, Vol 4, Iss 3, p 48 (2019) Biomimetics Volume 4 Issue 3 |
| ISSN: | 2313-7673 |
| Popis: | Mesenchymal stem cells (MSCs) are progenitors for bone-forming osteoblasts and lipid-storing adipocytes, two major lineages co-existing in bone marrow. When isolated in vitro, these stem cells recapitulate osteoblast or adipocyte formation if treated with specialised media, modelling how these lineages interact in vivo. Osteogenic differentiation is characterised by mineral deposits accumulating in the extracellular matrix, typically assessed using histological techniques. Adipogenesis occurs with accumulation of intracellular lipids that can be routinely visualised by Oil Red O staining. In both cases, staining requires cell fixation and is thus limited to end-point assessments. Here, a vital staining approach was developed to simultaneously detect mineral deposits and lipid droplets in differentiating cultures. Stem cells induced to differentiate produced mixed cultures containing adipocytes and bone-like nodules, and after two weeks live cultures were incubated with tetracycline hydrochloride and Bodipy to label mineral- and lipid-containing structures, respectively. Fluorescence microscopy showed the simultaneous visualisation of mineralised areas and lipid-filled adipocytes in live cultures. Combined with the nuclear stain Hoechst 33258, this approach further enabled live confocal imaging of adipogenic cells interspersed within the mineralised matrix. This multiplex labelling was repeated at subsequent time-points, demonstrating the potential of this new approach for the real-time high-precision imaging of live stem cells. |
| Databáze: | OpenAIRE |
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