Optimal labeling dose, labeling time, and magnetic resonance imaging detection limits of ultrasmall superparamagnetic iron-oxide nanoparticle labeled mesenchymal stromal cells

Anders Bruun Mathiasen, Louise Hansen, Tina Friis, Carsten Thomsen, Kishore Bhakoo, Jens Kastrup, Anders Bruun Mathiasen, Louise Hansen, Tina Friis, Carsten Thomsen, Kishore Bhakoo, Jens Kastrup

Abstract

Background. Regenerative therapy is an emerging treatment modality. To determine migration and retention of implanted cells, it is crucial to develop noninvasive tracking methods. The aim was to determine ex vivo magnetic resonance imaging (MRI) detection limits of ultrasmall superparamagnetic iron-oxide (USPIO) labeled mesenchymal stromal cells (MSCs). Materials and Methods. 248 gel-phantoms were constructed and scanned on a 1.5T MRI-scanner. Phantoms contained human MSCs preincubated with USPIO nanoparticles for 2, 6, or 21 hours using 5 or 10 μ g USPIO/10(5) MSCs. In addition, porcine hearts were scanned after injection of USPIO labeled MSCs. Results. Using 21 h incubation time and 10 μ g USPIO/10(5) MSCs, labeled cells were clearly separated from unlabeled cells on MRI using 250.000 (P < 0.001), 500.000 (P = 0.007), and 1.000.000 MSCs (P = 0.008). At lower incubation times and doses, neither labeled nor unlabeled cells could be separated. In porcine hearts labeled, but not unlabeled, MSCs were identified on MRI. Conclusions. As few as 250.000 MSCs can be detected on MRI using 21 h incubation time and 10 μ g USPIO/10(5) MSCs. At lower incubation times and doses, several million cells are needed for MRI detection. USPIO labeled cells can be visualized by MRI in porcine myocardial tissue.

Figures

Figure 1
Figure 1
MRI phantoms. (a) Two phantoms containing USPIO labeled cells. (b) MRI image of 2 phantoms with an ellipsoid region of interest placed in the upper phantom. USPIO: ultrasmall superparamagnetic iron-oxide.
Figure 2
Figure 2
Cellular iron content. The iron content per cell was determined in unlabeled MSC and MSC incubated with half or full dose USPIO for 2, 6, and 21 hours. MSC: mesenchymal stromal cells. USPIO: ultrasmall superparamagnetic iron-oxide. USPIO dose—full: 10 μg per 105 cells; half: 5 μg per 105 cells.
Figure 3
Figure 3
Absolute phantom MRI intensities after 21 hour USPIO incubation. MRI intensities are absolute mean values. USPIO: ultrasmall superparamagnetic iron-oxide. USPIO dose: full = 10 μg per 105 cells; half = 5 μg per 105 cells.
Figure 4
Figure 4
Phantoms intensity differences after 21-hour USPIO incubation. MRI intensity differences are the mean numeric difference between absolute MRI intensities of phantoms and reference gels. USPIO = ultrasmall superparamagnetic iron-oxide. USPIO dose: full = 10 μg per 105 cells; half = 5 μg per 105 cells.
Figure 5
Figure 5
MRI images of porcine myocardium before and after USPIO labeled MSC injection. T2*-images of porcine myocardium before injection (a1–a5) and after injection (b1–b5) of USPIO-labeled MSCs. USPIO labeled MSCs are identified as hypointense areas (arrows). MSCs: mesenchymal stromal cells. USPIO: ultrasmall superparamagnetic iron-oxide.
Figure 6
Figure 6
MRI images of porcine myocardium before and after unlabeled MSC injection. T2*-images of porcine myocardium before injection (a1–a5) and after injection (b1–b5) of unlabeled MSCs. Unlabeled cells cannot be identified. MSCs: mesenchymal stromal cells. USPIO: ultrasmall superparamagnetic iron-oxide.

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