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Methodological aspects of MRI of transplanted superparamagnetic iron oxide-labeled mesenchymal stem cells in live rat brain.

ABSTRACT: In vivo tracking of transplanted mesenchymal stem cells (MSCs) migration and homing is vital for understanding the mechanisms of beneficial effects of MSCs transplantation in animal models of diseases and in clinical trials. Transplanted cells can be labeled with superparamagnetic iron oxide (SPIO) particles and visualized in vivo using a number of iron sensitive MRI techniques. However, the applicability of those techniques for SPIO-labeled MSCs tracking in live brain has not been sufficiently investigated. The goal of this study was to estimate the efficiency of various MRI techniques of SPIO-labeled cell tracing in the brain. To achieve that goal, the precision and specificity of T2WI, T2*WI and SWI (Susceptibility-Weighted Imaging) techniques of SPIO-labeled MSCs tracing in vitro and in live rat brain were for the first time compared in the same experiment. We have shown that SWI presents the most sensitive pulse sequence for SPIO-labeled MSCs MR visualization. After intracerebral administration due to limitations caused by local micro-hemorrhages the visualization threshold was 102 cells, while after intra-arterial transplantation SWI permitted detection of several cells or even single cells. There is just one publication claiming detection of individual SPIO-labeled MSCs in live brain, while the other state much lower sensitivity, describe detection of different cell types or high resolution tracing of MSCs in other tissues. This study confirms the possibility of single cell tracing in live brain and outlines the necessary conditions. SWI is a method convenient for the detection of single SPIO labeled MSCs and small groups of SPIO labeled MSCs in brain tissue and can be appropriate for monitoring migration and homing of transplanted cells in basic and translational neuroscience.

SUBMITTER: Namestnikova D

PROVIDER: S-EPMC5648235 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Endothelial progenitor cells (EPCs) have been discovered to be relevant to the prognosis of cardiovascular diseases. Previous research has demonstrated that EPCs serve vital roles in the occurrence and development of atherosclerosis. Significant improvements have been made in MRI technology and in the experimental use of EPCs for therapeutic angiogenesis and vascular repair. Nevertheless, the migratory, adhesive, proliferative and angiogenic properties of EPCs remain unknown. The aims of the present study were to investigate the potential of using non?invasive monitoring with ultrasmall superparamagnetic iron oxide nanoparticle (USPION)?labeled endothelial progenitor cells (EPCs) after transplantation, and to assess the treatment outcomes in an atherosclerotic rabbit model. EPCs derived from rabbit peripheral blood samples were labeled with USPION?poly?l?lysine (USPION?PLL). The morphology, proliferation, adhesive ability and labeling efficiency of the EPCs were determined by optical and electron microscopy. Moreover, biological activity was assessed by flow cytometry. In addition, T2?weighted image fast spin?echo MRI was used to detect cell labeling. USPION content in the labeled EPCs was determined by Prussian blue staining and scanning electron microscopy. Rabbit atherosclerosis model was established using a high?fat diet. USPION?labeled EPCs were transplanted into rabbits, and in vivo MRI was performed 1 and 7 days after transplantation. It was found that EPCs cultured on Matrigel formed capillary?like structures, and expressed the surface markers CD133, CD31, CD34 and vascular endothelial growth factor receptor 2 (VEGFR2). The optimal USPION concentration was 32 µg/ml, as determined by adhesion and proliferation assays. It was identified that USPION?PLL nanoparticles were 10?20 nm in diameter. Histopathological analysis results indicated that 1 day after transplantation of the labeled EPCs, blue?stained granules were observed in the intima of vascular lesions in rabbit models after Prussian blue staining. Therefore, the present results suggest that USPION?labeled EPCs may play a role in repairing endothelial injury and preventing atherosclerosis in vivo.

Project description:BackgroundThe aim of this study was to develop an antiGPC3-ultrasuperparamagnetic iron oxide (USPIO) probe for early detection of hepatocellular carcinoma.MethodsGPC3 and AFP receptors were selected as biomarkers and conjugated with USPIO nanoparticles coated by dextran with carboxylate groups to synthesize antiGPC3-USPIO and antiAFP-USPIO probes. HepG2 cells (a human hepatocellular carcinoma cell model with high expression of GPC3) were used along with SMMC-7721 cells (a hepatocellular carcinoma cell model with no expression of GPC3), HeLa cells (a cervical cancer model), and HL-7702 (normal hepatocytes) which were used as controls. After incubation with the probes, the iron content in the cells was calculated, USPIO nanoparticles in cells were observed using transmission electron microscopy, and T1 and T2 relaxation times were measured with a 1.5 T magnetic resonance scanner.ResultsAntiGPC3-USPIO probes with a mean hydrodynamic diameter of 47 nm showed good biological compatibility. Transmission electron microscopic images indicated that the amount of USPIO nanoparticles taken up was significantly higher in HepG2 cells incubated with antiGPC3-USPIO than that in HepG2 cells incubated with antiAFP-USPIO or USPIO nanoparticles and that in the SMMC-7721 or HeLa cells incubated with antiGPC3-USPIO probes, antiAFP-USPIO probes, or USPIO nanoparticles. The higher the concentration and the longer the incubation time, the greater the number of USPIO nanoparticles found in the cells. No USPIO nanoparticles were found in the HL-7702 cells. All of the HepG2, SMMC-7721, and HeLa cells incubated with antiGPC3-USPIO, antiAFP-USPIO, or USPIO nanoparticles were able to shorten the T1 and T2 values in agar solution, especially the T2 images of HepG2 cells incubated with antiGPC3-USPIO probes.ConclusionAntiGPC3-USPIO probes can be utilized as a specific magnetic resonance targeting contrast agent for early detection of hepatocellular carcinoma. Using a 1.5 T magnetic resonance scanner, the optimal time for imaging HepG2 cells was around 2-4 hours after incubation with antiGPC3-USPIO probes.

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Methodological aspects of MRI of transplanted superparamagnetic iron oxide-labeled mesenchymal stem cells in live rat brain.

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