Abstract
This study reports the use of gold nanoparticle-based surface-enhanced Raman scattering (SERS) for probing the differentiation of mouse embryonic stem (mES) cells, including undifferentiated single cells, embryoid bodies (EBs), and terminally differentiated cardiomyocytes. Gold nanoparticles (GNPs) were successfully delivered into all 3 mES cell differentiation stages without affecting cell viability or proliferation. Transmission electron microscopy (TEM) confirmed the localization of GNPs inside the following cell organelles: mitochondria, secondary lysosome, and endoplasmic reticulum. Using bright- and dark-field imaging, the bright scattering of GNPs and nanoaggregates in all 3 ES cell differentiation stages could be visualized. EB (an early differentiation stage) and terminally differentiated cardiomyocytes both showed SERS peaks specific to metabolic activity in the mitochondria and to protein translation (amide I, amide II, and amide III peaks). These peaks have been rarely identified in undifferentiated single ES cells. Spatiotemporal changes observed in the SERS spectra from terminally differentiated cardiomyocyte tissues revealed local and dynamic molecular interactions as well as transformations during ES cell differentiation.
Highlights
Embryonic stem (ES) cells are pluripotent cells that have the capability to self-renew and differentiate into multiple tissue types
Ultramicroscopic imaging of Gold nanoparticles (GNPs) accumulation Nanoparticle entry and localization inside ES cells, embryoid bodies (EBs), and cardiomyocyte tissues were confirmed by Transmission electron microscopy (TEM)
GNPs were localized in the perinuclear region and in cell organelles such as the mitochondria, secondary lysosomes, and rough endoplasmic reticulum but not in the nucleus (Figures 3A– C)
Summary
Embryonic stem (ES) cells are pluripotent cells that have the capability to self-renew and differentiate into multiple tissue types. Identification of markers specific to each differentiation stage is essential for tracking the differentiation of ES cells Techniques such as immunocytochemistry, fluorescence microscopy, polymerase chain reaction, and RNA in situ hybridization are generally used to measure the expression of stage-specific embryonic antigen-1, and POU family transcription factors Oct4/Oct-3 [8], and CD9 [9] for undifferentiated state of ES cells. These techniques have certain limitations: they involve lengthy procedures lasting hours or days; require a large number of cells, labels, or markers; and cannot be carried out on living cells as they involve lysis, fixation, or both. In this rapidly expanding field, the need for faster noninvasive methods to characterize and monitor the differentiation of ES cells in situ and in real-time is more evident than ever before
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