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Open AccessArticle

Live Simultaneous Monitoring of Mineral Deposition and Lipid Accumulation in Differentiating Stem Cells

1
Wolfson STEM Centre, School of Medicine, The University of Nottingham, Nottingham NG7 2RD, UK
2
School of Life Sciences, University of Nottingham, Nottingham NG7 2RD, UK
3
The Early Life Research Unit, Division of Child Health, School of Medicine, The University of Nottingham, Nottingham NG7 2RD, UK
4
Advanced Materials Group, Faculty of Engineering, The University of Nottingham, Nottingham NG7 2RD, UK
5
Arthritis Research UK Pain Centre, The University of Nottingham, Nottingham NG7 2RD, UK
*
Author to whom correspondence should be addressed.
These authors contributed equally to the study.
Biomimetics 2019, 4(3), 48; https://doi.org/10.3390/biomimetics4030048
Received: 3 May 2019 / Revised: 22 June 2019 / Accepted: 4 July 2019 / Published: 10 July 2019
(This article belongs to the Special Issue Selected Papers from N.I.C.E. 2018)
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. View Full-Text
Keywords: live monitoring; stem cell; mineralisation; lipid detection live monitoring; stem cell; mineralisation; lipid detection
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MDPI and ACS Style

De Melo, N.; McGinlay, S.; Markus, R.; Macri-Pellizzeri, L.; Symonds, M.E.; Ahmed, I.; Sottile, V. Live Simultaneous Monitoring of Mineral Deposition and Lipid Accumulation in Differentiating Stem Cells. Biomimetics 2019, 4, 48.

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