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Label-free assessment of adipose-derived stem cell differentiation using coherent anti-Stokes Raman scattering and multiphoton microscopy.

TitleLabel-free assessment of adipose-derived stem cell differentiation using coherent anti-Stokes Raman scattering and multiphoton microscopy.
Publication TypeJournal Article
Year of Publication2012
AuthorsMouras R, Bagnaninchi PO, Downes AR, Elfick APD
JournalJ Biomed Opt
Volume17
Issue11
Pagination116011
Date Published2012 Nov
ISSN1560-2281
KeywordsAdipocytes, Adipose Tissue, Adult Stem Cells, Cell Differentiation, Cells, Cultured, Extracellular Matrix, Humans, Lipid Metabolism, Microscopy, Fluorescence, Multiphoton, Optical Phenomena, Osteoblasts, Spectrum Analysis, Raman
Abstract

Adult stem cells (SCs) hold great potential as likely candidates for disease therapy but also as sources of differentiated human cells in vitro models of disease. In both cases, the label-free assessment of SC differentiation state is highly desirable, either as a quality-control technology ensuring cells to be used clinically are of the desired lineage or to facilitate in vitro time-course studies of cell differentiation. We investigate the potential of nonlinear optical microscopy as a minimally invasive technology to monitor the differentiation of adipose-derived stem cells (ADSCs) into adipocytes and osteoblasts. The induction of ADSCs toward these two different cell lineages was monitored simultaneously using coherent anti-Stokes Raman scattering, two photon excitation fluorescence (TPEF), and second harmonic generation at different time points. Changes in the cell's morphology, together with the appearance of biochemical markers of cell maturity were observed, such as lipid droplet accumulation for adipo-induced cells and the formation of extra-cellular matrix for osteo-induced cells. In addition, TPEF of flavoproteins was identified as a proxy for changes in cell metabolism that occurred throughout ADSC differentiation toward both osteoblasts and adipocytes. These results indicate that multimodal microscopy has significant potential as an enabling technology for the label-free investigation of SC differentiation.

DOI10.1117/1.JBO.17.11.116011
Alternate JournalJ Biomed Opt
PubMed ID23117806
Grant ListG0802632 / / Medical Research Council / United Kingdom
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