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WIREs Nanomed Nanobiotechnol
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Cell tracking using gold nanoparticles and computed tomography imaging

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Cell‐based therapies utilize transplantation of living cells with therapeutic traits to alleviate numerous diseases and disorders. The use of such biological agents is an attractive alternative for diseases that existing medicine cannot effectively treat. Although very promising, translating cell therapy to the clinic has proven to be challenging, due to inconsistent results in preclinical and clinical studies. To examine the underlying cause for these inconsistencies, it is crucial to noninvasively monitor the accuracy of cell injection, and cell survival and migration patterns. The combination of classical imaging techniques with cellular contrast agents—mainly nanotechnological‐based—has enabled significant developments in cell‐tracking methodologies. One novel methodology, based on computed tomography (CT) as an imaging modality and gold nanoparticles (AuNPs) as contrast agents, has recently gained interest for its clinical applicability and cost‐effectiveness. Studies have shown that AuNPs can be used to efficiently label a variety of cell types, including stem cells and immune cells, without damaging their therapeutic efficacy. Successful in vivo experiments have demonstrated noninvasive, quantitative and longitudinal cell tracking with high sensitivity. This concept has the potential to be used not only as a research tool, but in clinical settings as well. WIREs Nanomed Nanobiotechnol 2018, 10:e1480. doi: 10.1002/wnan.1480 This article is categorized under: Diagnostic Tools > In Vivo Nanodiagnostics and Imaging
Microscopy images of cells post labeling with gold nanoparticles (AuNPs). Top: Intracellular uptake of AuNP complexed with poly‐l‐lysine and rhodamine B isothiocyanate (RITC) in human mesenchymal stem cells (MSCs). Shown are (a) bright field, (b) fluorescence (blue = DAPI, red = RITC), and (c) TEM images. Center: (a) SEM image of a labeled C6 cancer cell, and b) zoom in of a cluster of AuNPs inside the cell. Bottom: (a)–(c) Dark field microscopy of A‐431 cancer cell line (blue) labeled with increasing concentrations of AuNPs. (Top: Reprinted with permission from Ref 26. Copyright 2017 John Wiley and Sons. Center: Reprinted with permission from Ref 61. Copyright 2013 Royal Society of Chemistry. Bottom: Reprinted with permission from Ref 22. Copyright 2015 Nature Publishing Group)
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Designing of gold nanoparticles of different sizes and coatings for cell labeling. Size, as well as coating of gold nanoparticles (AuNPs), affects their cell labeling abilities. Left: Transmission electron microscopy (TEM) images of spherical AuNPs of increasing size, from 15 to 150 nm. Right: Schematic depiction of the range of AuNP sizes used in the study, and the chemical structures of the ligands used as coatings. Ligands examined represent different functionalities and charges. (Reprinted with permission from Ref . Copyright 2017 American Chemical Society)
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Noninvasive cell tracking by gold nanoparticle (AuNP) labeling. The cells are first labeled with AuNPs in vitro, then intravenously injected to the subject. In vivo computed tomography (CT) imaging enables real time, noninvasive cell tracking.
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Cell tracking with simultaneous monitoring of muscle recovery. Imaging cell treatment of muscle in a mouse model for Duchenne muscular dystrophy (MDX). (a) 3D volume‐rendered computed tomography (CT) scan of mouse 2 weeks posttransplantation of AuNP‐loaded mesenchymal stem cells in the right limb (arrow indicates injection site). Yellow: AuNPs located at the injection site, blue: calcification. Calcification is considerably more pronounced in the untreated left limb as compared to the treated limb. (b) 2D cross‐sectional slice. (c) Untreated MDX mouse; calcification signal is observed in both limbs. (Reprinted with permission from Ref . Copyright 2017 Elsevier)
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3D in vivo volume rendering micro‐computed tomography (CT) scans of brain post injection of AuNP‐labeled hMSC into the left ventricle. (a) One‐hour post injection; (b) 24 h post injection; (c) 1 month post injection; (d) 1 month post free AuNP injection (control rat). (Reprinted with permission from Ref . Copyright 2014 American Chemical Society)
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Computed tomography (CT) scans demonstrating migration of gold nanoparticle (AuNP)‐labeled T‐cells and their whole‐body biodistribution. (a) 3D volume rendering CT image of T‐cells that accumulated in the lungs 48 h post injection. Yellow areas represent AuNP‐labeled T‐cells. (b) Representative 2D CT image of lungs. Arrow indicates gold‐labeled cells. (c) Maximum intensity projection of micro‐CT scans 48 h post injection. Circles demarcate T‐cell accumulation in the tumor area. (Reprinted with permission from Ref . Copyright 2015 American Chemical Society)
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The key to efficient cell labeling. Achieving a balance between high nanoparticle uptake for maximum contrast while concurrently maintaining cell viability and function. This balance is controlled by five different factors: Particle size, shape and coating, loading time and concentration.
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Cell proliferation and functionality assays for T‐cells. (a) Proliferation assay with carboxyfluorescein succinimidyl ester (CFSE). CFSE‐labeled T‐cells loaded with increasing AuNP concentrations (0.35, 0.55, 0.70, and 1 mg/mL), stimulated for 3 days and analyzed for CFSE dilution. Data shown as average percentage of proliferative cells, normalized to control [cells without AuNPs (W/O)] ± SEM. No significant differences were observed between cells loaded with the different amounts of AuNPs and controls (p > 0.05). (b) Functionality assay. T‐cells loaded with increasing AuNP concentrations (0.35, 0.55, 0.70, and 1 mg/mL), co‐cultured with a positive target tumor cell line (888‐A2) for 120 min. IFN‐γ secretion (measured by ELISA) was normalized to control [cells w/o AuNPs (W/O)] ± SEM (n = 3) (p < 0.05, Student's paired t‐test). Cell‐function was impaired only for high gold concentrations (0.75, 1 mg/mL), after 120 min of incubation. (Reprinted with permission from Ref . Copyright 2015 Nature Publishing Group)
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