ABSTRACT
PURPOSE: 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. PROCEDURES: 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. RESULTS: 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). CONCLUSION: 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.
Subject(s)
Contrast Media , Fluorescent Dyes , Mesenchymal Stem Cells/cytology , Cell Differentiation , Cells, Cultured , Dextrans , Glycosaminoglycans/metabolism , Humans , Magnetic Resonance Imaging , Magnetite Nanoparticles , Mesenchymal Stem Cells/metabolism , Metal NanoparticlesABSTRACT
Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) have demonstrated the ability to improve myocardial function following transplantation into an ischemic heart; however, the functional benefits are transient possibly due to poor cell retention. A diagnostic technique that could visualize transplanted hESC-CMs could help to optimize stem cell delivery techniques. Thus, the purpose of this study was to develop a labeling technique for hESCs and hESC-CMs with the FDA-approved contrast agent indocyanine green (ICG) for optical imaging (OI). hESCs were labeled with 0.5, 1.0, 2.0, and 2.5 mg/ml of ICG for 30, 45, and 60 min at 37 degrees C. Longitudinal OI studies were performed with both hESCs and hESC-CMs. The expression of surface proteins was assessed with immunofluorescent staining. hESCs labeled with 2 mg ICG/ml for 60 min achieved maximum fluorescence. Longitudinal studies revealed that the fluorescent signal was equivalent to controls at 120 h postlabeling. The fluorescence signal of hESCs and hESC-CMs at 1, 24, and 48 h was significantly higher compared to precontrast data (p < 0.05). Immunocytochemistry revealed retention of cell-specific surface and nuclear markers postlabeling. These data demonstrate that hESCs and hESC-CMs labeled with ICG show a significant fluorescence up to 48 h and can be visualized with OI. The labeling procedure does not impair the viability or functional integrity of the cells. The technique may be useful for assessing different delivery routes in order to improve the engraftment of transplanted hESC-CMs or other stem cell progenitors.
Subject(s)
Embryonic Stem Cells/cytology , Fluorescent Antibody Technique/methods , Indocyanine Green/pharmacology , Myocytes, Cardiac/cytology , Staining and Labeling/methods , Stem Cell Transplantation/methods , Biomarkers/analysis , Biomarkers/metabolism , Cell Culture Techniques/methods , Cell Survival/drug effects , Cell Survival/physiology , Cells, Cultured , Embryonic Stem Cells/drug effects , Embryonic Stem Cells/physiology , Fluorescence , Heart Diseases/surgery , Humans , Membrane Proteins/analysis , Membrane Proteins/metabolism , Microscopy, Fluorescence/methods , Myocytes, Cardiac/drug effects , Myocytes, Cardiac/physiology , Time FactorsABSTRACT
PURPOSE: Peri-tumoral inflammation is a common tumor response that plays a central role in tumor invasion and metastasis, and inflammatory cell recruitment is essential to this process. The purpose of this study was to determine whether injected fluorescently-labeled monocytes accumulate within murine breast tumors and are visible with optical imaging. MATERIALS AND METHODS: Murine monocytes were labeled with the fluorescent dye DiD and subsequently injected intravenously into 6 transgenic MMTV-PymT tumor-bearing mice and 6 FVB/n control mice without tumors. Optical imaging (OI) was performed before and after cell injection. Ratios of post-injection to pre-injection fluorescent signal intensity of the tumors (MMTV-PymT mice) and mammary tissue (FVB/n controls) were calculated and statistically compared. RESULTS: MMTV-PymT breast tumors had an average post/pre signal intensity ratio of 1.8+/- 0.2 (range 1.1-2.7). Control mammary tissue had an average post/pre signal intensity ratio of 1.1 +/- 0.1 (range, 0.4 to 1.4). The p-value for the difference between the ratios was less than 0.05. Confocal fluorescence microscopy confirmed the presence of DiD-labeled cells within the breast tumors. CONCLUSION: Murine monocytes accumulate at the site of breast cancer development in this transgenic model, providing evidence that peri-tumoral inflammatory cell recruitment can be evaluated non-invasively using optical imaging.
Subject(s)
Breast Neoplasms/pathology , Inflammation/pathology , Mammary Neoplasms, Experimental/pathology , Microscopy, Fluorescence/methods , Animals , Breast Neoplasms/immunology , Female , Fluorescent Dyes/metabolism , Humans , Mammary Neoplasms, Experimental/immunology , Mammary Tumor Virus, Mouse/genetics , Mice , Mice, Transgenic , Microscopy, Confocal/methods , Monocytes/cytology , Monocytes/metabolismABSTRACT
The objective of this work is to establish an optical imaging technique that would enable monitoring of the integration of mesenchymal stem cells (MSC) in arthritic joints. Our approach is based on first developing a labeling technique of MSC with the fluorescent dye DiD followed by tracking the cell migration kinetics from the spatial distribution of the DiD fluorescence in optical images (OI). The experimental approach involves first the in vitro OI of MSC labeled with DiD accompanied by fluorescence microscopy measurements to establish localization of the signal within the cells. Thereafter, DiD-labeled MSC were injected into polyarthritic, athymic rats and the signal localization within the experimental animals was monitored over several days. The experimental results indicate that DiD integrated into the cell membrane. DiD-labeled MSC localization in the arthritic ankle joints was observed with OI indicating that this method can be applied to monitor MSC in arthritic joints.
Subject(s)
Arthritis/immunology , Arthritis/pathology , Image Enhancement/methods , Image Interpretation, Computer-Assisted/methods , Mesenchymal Stem Cells/immunology , Mesenchymal Stem Cells/pathology , Animals , Cell Movement/immunology , Cells, Cultured , Disease Models, Animal , Female , Humans , Microscopy, Fluorescence , Rats , Rats, NudeABSTRACT
OBJECTIVE: The objective of our study was to determine the frequency and cause of anterior layering of excreted 18F-FDG in the bladder on PET/CT. CONCLUSION: Anterior layering of excreted FDG in the bladder is commonly seen on PET/CT scans obtained with i.v. iodinated contrast material and is due to displacement of FDG by excreted iodinated contrast material; this phenomenon may unmask FDG-avid bladder disease.
