Noninvasive Detection and Imaging of Molecular Markers in Live Cardiomyocytes Derived from Human Embryonic Stem Cells

Raman microspectroscopy (RMS) was used to detect and image molecular markers specific to cardiomyocytes (CMs) derived from human embryonic stem cells (hESCs). This technique is noninvasive and thus can be used to discriminate individual live CMs within highly heterogeneous cell populations. Principa...

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Bibliographic Details
Published in:Biophysical journal 2011-01, Vol.100 (1), p.251-259
Main Authors: Pascut, Flavius C., Goh, Huey T., Welch, Nathan, Buttery, Lee D., Denning, Chris, Notingher, Ioan
Format: Article
Language:English
Subjects:
Algorithms
Animals
Biomarkers - metabolism
cardiomyocytes
Cell Survival
Cells
Comparative analysis
embryonic stem cells
Embryonic Stem Cells - cytology
genetic markers
Genotype & phenotype
glycogen
Human
Humans
image analysis
Imaging
Lasers
Markers
Mice
Molecular Imaging - methods
Myocytes, Cardiac - cytology
Myocytes, Cardiac - metabolism
Nonnative species
Phenotype
Populations
Principal Component Analysis
progeny
Proteins
Spectra
Spectroscopy, Imaging, and Other Techniques
Spectrum analysis
Spectrum Analysis, Raman
Stem cells
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Summary:Raman microspectroscopy (RMS) was used to detect and image molecular markers specific to cardiomyocytes (CMs) derived from human embryonic stem cells (hESCs). This technique is noninvasive and thus can be used to discriminate individual live CMs within highly heterogeneous cell populations. Principal component analysis (PCA) of the Raman spectra was used to build a classification model for identification of individual CMs. Retrospective immunostaining imaging was used as the gold standard for phenotypic identification of each cell. We were able to discriminate CMs from other phenotypes with >97% specificity and >96% sensitivity, as calculated with the use of cross-validation algorithms (target 100% specificity). A comparison between Raman spectral images corresponding to selected Raman bands identified by the PCA model and immunostaining of the same cells allowed assignment of the Raman spectral markers. We conclude that glycogen is responsible for the discrimination of CMs, whereas myofibril proteins have a lesser contribution. This study demonstrates the potential of RMS for allowing the noninvasive phenotypic identification of hESC progeny. With further development, such label-free optical techniques may enable the separation of high-purity cell populations with mature phenotypes, and provide repeated measurements to monitor time-dependent molecular changes in live hESCs during differentiation in vitro.
ISSN:0006-3495
1542-0086
DOI:10.1016/j.bpj.2010.11.043