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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Available from: Heike Daldrup-Link, Oct 09, 2015
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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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    ABSTRACT: Aim The aim of this work was the development of successful cell therapy techniques for cartilage engineering. This will depend on the ability to monitor non-invasively transplanted cells, especially mesenchymal stem cells (MSCs) that are promising candidates to regenerate damaged tissues. Methods MSCs were labeled with superparamagnetic iron oxide particles (SPIO). We examined the effects of long-term labeling, possible toxicological consequences and the possible influence of progressive concentrations of SPIO on chondrogenic differentiation capacity. Results No influence of various SPIO concentrations was noted on human bone marow MSC viability or proliferation. We demonstrated long-term (4 weeks) in vitro retention of SPIO by human bone marrow MSCs seeded in collagenic sponges under TGF-β1 chondrogenic conditions, detectable by Magnetic Resonance Imaging (MRI) and histology. Chondrogenic differentiation was demonstrated by molecular and histological analysis of labeled and unlabeled cells. Chondrogenic gene expression (COL2A2, ACAN, SOX9, COL10, COMP) was significantly altered in a dose-dependent manner in labeled cells, as were GAG and type II collagen staining. As expected, SPIO induced a dramatic decrease of MRI T2 values of sponges at 7T and 3T, even at low concentrations. Conclusions This study clearly demonstrates (1) long-term in vitro MSC traceability using SPIO and MRI and (2) a deleterious dose-dependence of SPIO on TGF-β1 driven chondrogenesis in collagen sponges. Low concentrations (12.5–25 µg Fe/mL) seem the best compromise to optimize both chondrogenesis and MRI labeling.
    PLoS ONE 05/2014; 9(5):e98451. DOI:10.1371/journal.pone.0098451 · 3.23 Impact Factor
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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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    ABSTRACT: Recent approaches of regenerative medicine can offer a therapeutic option for patients undergoing acute kidney injury. In particular, mesenchymal stem cells were shown to ameliorate renal function and recovery after acute damage. We here evaluated the protective effect and localization of CD133+ renal progenitors from the human inner medulla in a model of glycerol-induced acute tubular damage and we compared the results with those obtained with bone marrow-derived mesenchymal stem cells. We found that CD133+ progenitor cells promoted the recovery of renal function, preventing tubular cell necrosis and stimulating resident cell proliferation and survival, similar to mesenchymal stem cells. In addition, by optical imaging analysis, CD133+ progenitor cells accumulated within the renal tissue, and a reduced entrapment in lung, spleen, and liver was observed. Mesenchymal stem cells were detectable at similar levels in the renal tissue, but a higher signal was present in extrarenal organs. Both cell types produced several cytokines/growth factors, suggesting that a combination of different mediators is involved in their biological action. These results indicate that human CD133+ progenitor cells are renotropic and able to improve renal regeneration in acute kidney injury.
    05/2014; 2(5). DOI:10.14814/phy2.12009
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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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    ABSTRACT: Mesenchymal stem cells (MSCs) contribute to the recovery of tissue injury, providing a paracrine support. Cell-derived extracellular vesicles (EVs), carrying membrane and cytoplasmatic constituents of the cell of origin, have been described as a fundamental mechanism of intercellular communication. We previously demonstrated that EVs derived from human MSCs accelerated recovery following acute kidney injury (AKI) in vivo. The aim of the present study was to investigate the biodistribution and the renal localization of EVs in AKI. For this purpose, two methods for EV labeling suitable for in vivo tracking with optical imaging (OI), were employed using near infrared (NIR) dye (DiD): i) labeled EVs were generated by MSCs pre-incubated with NIR dye and collected from cell supernatants; ii) purified EVs were directly labeled with NIR dye. EVs obtained with these two procedures were injected intravenously (i.v.) into mice with glycerol-induced AKI and into healthy mice to compare the efficacy of the two labeling methods for in vivo detection of EVs at the site of damage. We found that the labeled EVs accumulated specifically in the kidneys of the mice with AKI compared with the healthy controls. After 5 h, the EVs were detectable in whole body images and in dissected kidneys by OI with both types of labeling procedures. The directly labeled EVs showed a higher and brighter fluorescence compared with the labeled EVs produced by cells. The signal generated by the directly labeled EVs was maintained in time, but provided a higher background than that of the labeled EVs produced by cells. The comparison of the two methods indicated that the latter displayed a greater specificity for the injured kidney.
    International Journal of Molecular Medicine 02/2014; 33(5). DOI:10.3892/ijmm.2014.1663 · 2.09 Impact Factor
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