Labeling Human Mesenchymal Stem Cells with Fluorescent Contrast Agents: the Biological Impact

Department of Radiology, University of California, San Francisco, San Francisco, CA, USA.
Molecular imaging and biology: MIB: the official publication of the Academy of Molecular Imaging (Impact Factor: 2.77). 04/2010; 13(1):3-9. DOI: 10.1007/s11307-010-0322-0
Source: PubMed


This study aims to determine the effect of human mesenchymal stem cell (hMSC) labeling with the fluorescent dye DiD and the iron oxide nanoparticle ferucarbotran on chondrogenesis.
hMSCs were labeled with DiD alone or with DiD and ferucarbotran (DiD/ferucarbotran). hMSCs underwent confocal microscopy, optical imaging (OI), and magnetic resonance (MR) imaging. Chondrogenesis was induced by transforming growth factor-b and confirmed by histopathology and glycosaminoglycan (GAG) production. Data of labeled and unlabeled hMSCs were compared with a t test.
Cellular uptake of DiD and ferucarbotran was confirmed with confocal microscopy. DiD labeling caused a significant fluorescence on OI, and ferucarbotran labeling caused a significant T2* effect on MR images. Compared to nonlabeled controls, progenies of labeled MSCs exhibited similar chondrocyte morphology after chondrogenic differentiation, but the labeled cells demonstrated significantly reduced GAG production (p < 0.05).
DiD and DiD/ferucarbotran labeling of hMSC does not interfere with cell viability or morphologic differentiation into chondrocytes, but labeled cells exhibit significantly less GAG production compared to unlabeled cells.

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    • "Discordant results were described by Heymer [31], working in collagen hydrogels at 1.5 mM, who reported no SPIO influence on chondrogenic gene expression, but a slightly more intense staining intensity in the pellets of unlabeled cells. In this way, Boddington [32] did not observe any influence of 100 µg Fe/mL on MSC-induced chondrogenesis (pellet) as assessed histologically, but found a significantly reduced GAG production in labeled cells. Other contradictory studies did not demonstrate any deleterious influence on MSC differentiation. "
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    • "The use of small-molecule fluorophores and, in particular, NIR molecules is a powerful tool to track stem cells for noninvasive visualization, giving the possibility to follow and to localize labeled cells in living whole-body animal. These dyes were employed for in vivo cell, antibody, and extracellular vesicle detection (Zou et al. 2009; Boddington et al. 2011; Hood et al. 2011). Using optical imaging, previous experiments showed the possibility to localize and compare the distribution of MSCs in the kidney and other organs of AKI animals after intravenous or intra-arterial injection (T€ ogel et al. 2008). "
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    • "Recently, the OI technique has been improved with the possibility of visualizing a few labeled cells in vivo by using new dyes (21–24). Good candidate dyes to maximize the depth of tissue penetration and reduce the background are near-infrared (NIR) fluorophores (700–900 nm); the absorption coefficient of tissue is very low and light possesses a high potential for penetration (25–27). "
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