Aptamer-based isolation and subsequent imaging of mesenchymal stem cells in ischemic myocard by magnetic resonance imaging.

PURPOSE: Mesenchymal stem cells (MSC) seem to be a promising cell source for cellular cardiomyoplasty. We recently developed a new aptamer-based specific selection of MSC to provide "ready to transplant" cells directly after isolation. We evaluated MRI tracking of newly isolated and freshly transplanted MSC in the heart using one short ex vivo selection step combining specific aptamer-based isolation and labeling of the cells. MATERIALS AND METHODS: Bone marrow (BM) was collected from healthy pigs. The animals were euthanized and the heart was placed in a perfusion model. During cold ischemia, immunomagnetic isolation of MSC from the BM by MSC-specific aptamers labeled with Dynabeads was performed within 2 h. For histological identification the cells were additionally stained with PKH26. Approx. 3 x 10(6) of the freshly aptamer-isolated cells were injected into the ramus interventricularis anterior (RIVA) and 5 x 10(5) cells were injected directly into myocardial tissue after damaging the respective area by freezing (cryo-scar). 3 x 10(6) of the aptamer-isolated cells were kept for further characterization (FACS and differentiation assays). 20 h after cell transplantation, MRI of the heart using a clinical 3.0 Tesla whole body scanner (Magnetom Trio, Siemens, Germany) was performed followed by histological examinations. RESULTS: The average yield of sorted cells from 120 ml BM was 7 x 10(6) cells. The cells were cultured and showed MSC-like properties. MRI showed reproducible artifacts within the RIVA-perfusion area and the cryo-scar with surprisingly excellent quality. The histological examination of the biopsies showed PKH26-positive cells within the areas which were positive in the MRI in contrast to the control biopsies. CONCLUSION: Immunomagnetic separation of MSC by specific aptamers linked to magnetic particles is feasible, effective and combines a specific separation and labeling technique to a "one stop shop" strategy.

Magnetic resonance imaging of iron-oxide labeled SK-Mel 28 human melanoma cells in the chick embryo using a clinical whole body MRI scanner.

PURPOSE: To evaluate advantages and limitations of magnetic resonance imaging (MRI) to monitor the migration of superparamagnetic iron oxide (SPIO) labeled cells in the chick embryo. MATERIALS AND METHODS: Labeled human SK-Mel 28 melanoma cells were injected into the E2 chick embryo neural tube. Embryos were examined with a clinical 3 T MRI whole body system using 3D T*(2)-weighted sequences with isotropic spatial resolutions of 0.3-1.0 mm. MR-measurements of embryos were performed 2 - 16 days after cell injection. MRI findings were verified by dissection and histology. RESULTS: After injection, melanoma cells formed aggregations that were detectable in the neural tube as signal voids in MR images from day 2 after injection. Emigrating cells later left MRI detectable tracks. Aggregates that remained in the neural tube left label that was absorbed by glia cells. In E18 chick embryos, signals of haematopoiesis interfered with signals from cell labeling. CONCLUSION: It was shown that SK-Mel 28 cells will resume the neural crest pathways after injection into the embryonic micro-environment. SPIO cell labeling allows monitoring of transplanted melanoma cells during embryonic development. MRI using the standard clinical equipment promises to be valuable for high-sensitive monitoring of ex-vivo labeled cells in the chick embryo.

Geometry and extension of signal voids in MR images induced by aggregations of magnetically labelled cells.

The ability of magnetic resonance imaging (MRI) to visualize magnetically labelled cells has attracted much attention for revealing cellular events. The present study addressed the geometry and the extension of signal voids in static signal dephasing MRI induced by aggregations of magnetically labelled cells by means of a three-dimensional numerical model. The magnetic field distortions around spherical cell aggregations were treated as equivalent to those of a magnetic dipole. Intravoxel signal dephasing and respective signal voids attributed to these field inhomogeneities were computed. Effects of cell concentration on the signal void in the plane of view were evaluated in terms of dipole magnetization. Signal void characteristics were scrutinized systematically for fundamental sequence parameters including echo time, voxel size and plane-of-view orientation. For all variables examined, significant changes in geometry as well as extension of signal voids were demonstrated. The results are of crucial importance to optimize and interpret MR images with regard to spatial accuracy as well as sensitivity to detect aggregations of labelled cells in vitro or even in vivo. It is anticipated that the dependence of the extension of signal voids on the local magnetization may be valuable for quantifying labelled cells.

Effect of concentration of SH U 555A labeled human melanoma cells on MR spin echo and gradient echo signal decay at 0.2, 1.5, and 3T.

In the current study the effect of increasing concentrations of superparamagnetic iron oxide labeled cells on the MRI signal decay at magnetic field strengths of 0.2, 1.5, and 3 T was evaluated. The spin echo and gradient echo cellular transverse relaxivity was systematically studied for various concentrations (N = 1, 5, 10, 20, 40, and 80 cells/microl(gel)) of homogeneously suspended SH U 555A labeled SK-Mel28 human melanoma cells. For all field strengths investigated a linear relationship between cellular transverse relaxation enhancement and cell concentration was found. In the spin echo case, the cellular relaxivities [i.e., d(deltaR2)/dN] were determined to 0.12 s(-1) (cell/microl)(-1) at 0.2 T, 0.16 s(-1) (cell/microl)(-1) at 1.5 T, and 0.17 s(-1) (cell/microl) at 3 T. In the gradient echo case, the calculated cellular relaxivities (i.e., d(deltaR2*)/dN) were 0.51 s(-1) (cell/microl)(-1) at 0.2 T, 0.69 s(-1) (cell/microl)(-1) at 1.5 T, and 0.71 s(-1) (cell/microl)(-1) at 3 T. The proposed preparation technique has proven to be a simple and reliable approach to quantify effects of magnetically labeled cells in vitro. On the basis of this quantification well suited tissue specific models can be derived.

Effect of spatial distribution of magnetic dipoles on Lamor frequency distribution and MR Signal decay--a numerical approach under static dephasing conditions.

Cells loaded with superparamagnetic iron oxide (SPIO) cause relatively strong magnetic field distortions, implying that field position effects of neighboring SPIO loaded cells have to be accounted for. We treated SPIO loaded cells as magnetic dipoles in a homogeneous magnetic field and computed the 3D frequency distribution and the related signal decay using a numerical approach under static dephasing conditions. The volume fraction of dipoles was kept constant for all simulations. For larger randomly distributed magnetic dipoles we found a non-Lorentzian frequency distribution and a non-monoexponential signal decay whereas, for smaller dipoles, the frequency distribution was more Lorentzian and the signal decay was well fitted monoexponentially. Moreover, based on our numerical and experimental findings, we found the gradient echo signal decay due to a single SPIO labeled cell to be non-monoexponential. The numerical approach provides deeper understanding of how the spatial distribution of SPIO loaded cells affects the MR signal decay. This fact has to be considered for the in vivo quantification of SPIO loaded cells, implying that in tissues with different spatial distributions of identical SPIO concentrations, different signal decays might be observed.

[A preparation technique for quantitative investigation of SPIO-containing solutions and SPIO-labelled cells by MRI]. / Eine Präparationstechnik zur quantitativen Untersuchung SPIO-haltiger Lösungen und SPIO-markierter Zellen in der MRT.

UNLABELLED: PURPOSE. This work aims to present a preparation technique for ex-vivo MR examination of SPIO (superparamagnetic iron oxide) containing solutions or SPIO labeled cells. Accumulations of SPIO particles and labeled cells were prepared in different concentrations using agar gel phantoms. Signal extinction around accumulations of magnetic material was examined systematically by gradient echo sequences with variable echo times and spatial resolution. The correlation between local iron concentration and diameter of signal extinction in MR gradient echo images was investigated. METHODS: Resovist, (SHU 555A) was used as superparamagnetic contrast medium. Different concentrations of SPIO-containing solutions (0.75 - 15 mg Fe/10 ml) and magnetically labeled SK-Mel28 cells (25,000-1,000,000 cells/10 ml) were accommodated inside a defined volume in an agar matrix. Diameters of signal void were assessed in dependence on local iron concentration, echo time (5-25 ms) and isotropic spatial resolution (length of voxel 0.25 - 0.60 mm). Measurements were performed on a clinical MR whole body scanner (3 Tesla) using a spoiled gradient echo sequence (FLASH). RESULTS: For the present experimental conditions sensitivity to detect the magnetic label was maximized using TE 25 ms. In contrast, the area of signal cancellation was minimized using TE 5 ms and isotropic resolution of 0.25 mm. In the latter case the image indicated the area of magnetic material most precisely. Diameter of signal cancellation was a logarithmic function on local iron concentration. In the presented set-up detection of concentrations as low as 0.75 mg Fe/10 ml in SPIO-containing solution or 1.25 mg Fe/10 ml in SPIO-labeled SK-Mel28 cells was certainly possible. CONCLUSION: The proposed preparation strategy with a well defined spatial distribution of the magnetic material in an agar gel phantom produced reliable results and appears clearly superior compared to set-ups with randomly distributed material in glass tubes. The diameter of the signal extinction in gradient echo images was significantly affected by the choice of echo time and spatial resolution. The calibration of signal cancellation versus iron concentrations may be valuable to assess SPIO concentrations and possibly numbers of labeled cells under specific conditions in vitro or even in vivo.