Abstract
The objectives of this study were to investigate the feasibility of imaging green fluorescent protein (GFP)-expressing cells labeled with iron oxide nanoparticles with the fast low-angle positive contrast steady-state free precession (FLAPS) method and to compare them with the traditional negative contrast technique. The GFP-R3230Ac cell line (GFP cell) was incubated for 24 hours using 20 microg Fe/mL concentration of superparamagnetic iron oxide (SPIO) and ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles. Cell samples were prepared for iron content analysis and cell function evaluation. The labeled cells were imaged using positive contrast with FLAPS imaging, and FLAPS images were compared with negative contrast T2*-weighted images. The results demonstrated that SPIO and USPIO labeling of GFP cells had no effect on cell function or GFP expression. Labeled cells were successfully imaged with both positive and negative contrast magnetic resonance imaging (MRI). The labeled cells were observed as a narrow band of signal enhancement surrounding signal voids in FLAPS images and were visible as signal voids in T2*-weighted images. Positive contrast and negative contrast imaging were both valuable for visualizing labeled GFP cells. MRI of labeled cells with GFP expression holds potential promise for monitoring the temporal and spatial migration of gene markers and cells, thereby enhancing the understanding of cell- and gene-based therapeutic strategies.
Highlights
A STANDARD METHOD FOR TRACKING a reporter gene within a cell of interest is to link it to a second gene encoding a fluorescent protein
The ability to image cells in vivo may be very useful for studying the effects of stem cell therapy, inflammation, tumors, immune response, etc
We demonstrated that iron oxide magnetic resonance contrast agents do not affect the marker gene expression within green fluorescent protein (GFP) cells, cell viability, or function
Summary
A STANDARD METHOD FOR TRACKING a reporter gene within a cell of interest is to link it to a second gene encoding a fluorescent protein. It has become possible to depict biologic processes at the cellular and molecular levels using magnetic resonance imaging (MRI).[2,3] Of primary interest is the development of MRI methods to noninvasively monitor the progress of targeted therapies.[3,4] It is highly. Dynamically monitoring therapeutic genes in tissues is challenging, and the distribution of the genes typically remains uncertain.[19] Real-time and direct MRI monitoring of genes within tissues is hindered by the target techniques.[20] The purpose of this study was (1) to label green fluorescent protein (GFP)expressing cells with USPIO and compare the labeling efficiency and GFP expression with previously reported SPIO-labeled GFP-expressing cells[21] and (2) to track the GFP-expressing cells using positive contrast methods and compare the resulting images with those obtained using more conventional negative contrast MRI techniques
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