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Labeling hESCs and hMSCs with Iron Oxide Nanoparticles for Non-Invasive in vivo Tracking with MR Imaging
Published on: March 31, 2008
In vitro imaging of human monocytic cellular activity using superparamagnetic iron oxide
Akihiro Nishie1, Kengo Yoshimitsu, Tomohiro Nakayama
1Department of Clinical Radiology, Graduate School of Medical Sciences, Kyushu University, 3-1-1, Maidashi, Higashi-ku Fukuoka, 812-8582, Japan. anishie@radiol.med.kyushu-u.ac.jp
Abstract:
The aim is to evaluate if a difference in activity of monocytic cells can be described on superparamagnetic iron oxide (SPIO)-magnetic resonance imaging (MRI) in vitro when a low concentration of SPIO is administered. Human monocytic cells were stimulated with phorbol 12-myristate 13-acetate (PMA) or left non-stimulated for 24 h and then treated with SPIO with a final concentration of 7.2 microg Fe/ml for 48 h. They were collected, and an MRI phantom was prepared by suspending them in 4% gelatin and scanned using gradient-echo T2*-weighted and spin-echo T2-weighted imaging with two different echo times. The signal intensity and T2*/T2 relaxation times of each layer were also assessed. The number and degree of intracellular filling of cells positively stained with Berlin blue were evaluated microscopically. In addition, the heme oxygenase-1 expression in non-stimulated monocytic cells or monocytic cells stimulated with PMA was evaluated using semiquantitative reverse transcription and polymerase chain reaction. The signal reduction of cells stimulated with PMA was 3.9- to 5.0-fold and 2.7- to 3.1-fold greater than that of non-stimulated cells on the T2*- and T2-weighted images, respectively. These signal reductions were also visually confirmed. Estimated T2*/T2 relaxation times of cells stimulated with PMA and non-stimulated cells were 31.3/137.8 and 122.8/182.6 ms, respectively. Cells positively stained with Berlin blue were observed in 4.5% and 25.3% of non-stimulated cells and those stimulated with PMA, respectively (p<0.05). The intracellular filling in cells stimulated with PMA was significantly larger than that in non-stimulated cells (p<0.05). The expression of the heme oxygenase-1 gene in monocytic cells stimulated with PMA was 2.9-fold greater than that in non-stimulated monocytic cells. SPIO-MRI can visually describe a difference in activity of monocytic cells in vitro when a low concentration of SPIO is administered.

