Research Papers: Imaging

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

[+] Author Affiliations
Rabah Mouras, Andrew R. Downes, Alistair P. D. Elfick

University of Edinburgh, School of Engineering, Institute for Materials and Processes, King’s Buildings, Mayfield Road, Edinburgh EH9 3JL, United Kingdom

Pierre O. Bagnaninchi

University of Edinburgh, MRC Centre for Regenerative Medicine, Edinburgh, EH16 4SB, United Kingdom

J. Biomed. Opt. 17(11), 116011 (Nov 01, 2012). doi:10.1117/1.JBO.17.11.116011
History: Received March 12, 2012; Revised September 27, 2012; Accepted September 28, 2012
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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.

Figures in this Article
© 2012 Society of Photo-Optical Instrumentation Engineers

Citation

Rabah Mouras ; Pierre O. Bagnaninchi ; Andrew R. Downes and Alistair P. D. Elfick
"Label-free assessment of adipose-derived stem cell differentiation using coherent anti-Stokes Raman scattering and multiphoton microscopy", J. Biomed. Opt. 17(11), 116011 (Nov 01, 2012). ; http://dx.doi.org/10.1117/1.JBO.17.11.116011


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