Magnetic resonance imaging of single co-labeled mesenchymal stromal cells after intracardial injection in mice.

PURPOSE: The aim of this study was to establish co-labeling of mesenchymal stromal cells (MSC) for the detection of single MSC in-vivo by MRI and histological validation. MATERIALS AND METHODS: Mouse MSC were co-labeled with fluorescent iron oxide micro-particles and carboxyfluorescein succinimidyl ester (CFSE). The cellular iron content was determined by atomic absorption spectrometry. Cell proliferation and expression of characteristic surface markers were determined by flow cytometry. The chondrogenic differentiation capacity was assessed. Different amounts of cells (n1 = 5000, n2 = 15 000, n3 = 50 000) were injected into the left heart ventricle of 12 mice. The animals underwent sequential MRI on a clinical 3.0 T scanner (Intera, Philips Medical Systems, Best, The Netherlands). For histological validation cryosections were examined by fluorescent microscopy. RESULTS: Magnetic and fluorescent labeling of MSC was established (mean cellular iron content 23.6 ± 3 pg). Flow cytometry showed similar cell proliferation and receptor expression of labeled and unlabeled MSC. Chondrogenic differentiation of labeled MSC was verified. After cell injection MRI revealed multiple signal voids in the brain and fewer signal voids in the kidneys. In the brain, an average of 4.6 ±â€Š1.2 (n1), 9.0 ±â€Š3.6 (n2) and 25.0 ± 1.0 (n3) signal voids were detected per MRI slice. An average of 8.7 ±â€Š3.1 (n1), 22.0 ±â€Š6.1 (n2) and 89.8 ±â€Š6.5 (n3) labeled cells per corresponding stack of adjacent cryosections could be detected in the brain. Statistical correlation of the numbers of MRI signal voids in the brain and single MSC found by histology revealed a correlation coefficient of r = 0.91. CONCLUSION: The study demonstrates efficient magnetic and fluorescent co-labeling of MSC and their detection on a single cell level in mice by in-vivo MRI and histology. The described techniques may broaden the methods for in-vivo tracking of MSC. KEY POINTS: • Detection of single magnetically labeled MSC in-vivo using a clinical 3.0 T MRI is possible.• Fluorescent and magnetic co-labeling does not affect cell vitality.• The number of cells detected by MRI and histology has a high correlation.

Superparamagnetic iron oxide nanoparticles in biomedicine: applications and developments in diagnostics and therapy.

Superparamagnetic iron oxide nanoparticles (SPIO) can be used to image physiological processes and anatomical, cellular and molecular changes in diseases. The clinical applications range from the imaging of tumors and metastases in the liver, spleen and bone marrow, the imaging of lymph nodes and the CNS, MRA and perfusion imaging to atherosclerotic plaque and thrombosis imaging. New experimental approaches in molecular imaging describe undirected SPIO trapping (passive targeting) in inflammation, tumors and associated macrophages as well as the directed accumulation of SPIO ligands (active targeting) in tumor endothelia and tumor cells, areas of apoptosis, infarction, inflammation and degeneration in cardiovascular and neurological diseases, in atherosclerotic plaques or thrombi. The labeling of stem or immune cells allows the visualization of cell therapies or transplant rejections. The coupling of SPIO to ligands, radio- and/or chemotherapeutics, embedding in carrier systems or activatable smart sensor probes and their externally controlled focusing (physical targeting) enable molecular tumor therapies or the imaging of metabolic and enzymatic processes. Monodisperse SPIO with defined physicochemical and pharmacodynamic properties may improve SPIO-based MRI in the future and as targeted probes in diagnostic magnetic resonance (DMR) using chip-based µNMR may significantly expand the spectrum of in vitro analysis methods for biomarker, pathogens and tumor cells. Magnetic particle imaging (MPI) as a new imaging modality offers new applications for SPIO in cardiovascular, oncological, cellular and molecular diagnostics and therapy.

[Magnetic resonance imaging of single SPIO labeled mesenchymal stem cells at 3 Tesla]. / Magnetresonanz-Bildgebung von einzelnen SPIO-markierten mesenchymalen Stammzellen bei 3 Tesla.

PURPOSE: To assess the detectability of single magnetically labeled mesenchymal stem cells (MSC) in-vitro on a clinical 3T MR scanner using a small animal volume coil. MATERIALS AND METHODS: GFP-transfected MSC were magnetically labeled with superparamagnetic iron oxide particles (SPIO) while applying different dosages of iron (56 vs. 560 microg Fe/ml). The cellular iron content was determined with atomic absorption spectrometry (AAS). Single labeled MSC were displayed in a culture flask using MR imaging and microscopy. Special cell phantoms were designed to examine the detection of labeled MSC with MR imaging in a spatial model. A T2*-weighted 3D gradient echo sequence with isotropic spatial resolution of 150 to 500 microm (3) was used for image acquisition. The detection of labeled MSC in the cell phantoms was quantitatively evaluated using an automated image analysis. Statistical analysis was performed with a significance level of p < 0.05. RESULTS: The labeling of MSC yielded a mean cellular iron content of 1.5 +/- 0.17 pg Fe/cell (56 microg Fe/ml) and 8.3 +/- 1.85 pg Fe/cell (560 microg Fe/ml). Examination of the culture flasks showed single magnetically labeled MSC centered in much larger MR signal voids. The detection and quantification of single MSC in cell phantoms were feasible for spatial resolutions of 150 microm and 200 microm. Cells with a lower SPIO content (1.5 +/- 0.17 pg Fe/cell) were detected in 14.2 +/- 4.2 % (150 microm) and 7.7 +/- 3.8 % (200 microm). MSC with a higher cellular SPIO content (8.3 +/- 1.85 pg Fe/cell) revealed significantly higher occurrences at both spatial resolutions with 81.4 +/- 5.8 % (150 microm) and 59.9 +/- 12.4 % (200 microm), respectively. Regarding the spatial resolution (150 vs. 200 microm), significantly different detection rates were determined only for MSC with the higher SPIO content (8.3 +/- 1.85 pg Fe/cell). CONCLUSION: Detection of single magnetically labeled MSC is feasible on a clinical 3T MR scanner with a small animal volume coil at isotropic spatial resolutions of 150 microm and 200 microm. The number of detected cells is influenced by the cellular iron content and the spatial resolution.