In Vivo Detection of Embryonic Stem Cell–Derived Cardiovascular Progenitor Cells Using Cy3-Labeled Gadofluorine M in Murine Myocardium

Abstract

Objective The aim of the current study is to test the ability to label and detect murine embryonic stem cell-derived cardiovascular progenitor cells (ES-CPCs) with Magnetic Resonance Imaging (MRI) using the novel contrast agent Gadofluorine M-Cy3 (GdFM-Cy3). Background Cell therapy shows great promise for the treatment of cardiovascular disease. An important limitation to previous clinical studies is the inability to accurately identify transplanted cells. GdFM-Cy3 is a lipophilic paramagnetic contrast agent that contains a perfluorinated side chain and an amphiphilic character that allows for micelle formation in an aqueous solution. Previous studies reported that it is easily taken up and stored within the cytosol of mesenchymal stem cells, thereby allowing for paramagnetic cell labeling. Investigators in our laboratory have recently developed techniques for the robust generation of ES-CPCs. We reasoned that GdFM-Cy3 would be a promising agent for the in vivo detection of these cells after cardiac cell transplantation. Methods ES-CPCs were labeled with GdFM-Cy3 by incubation. In vitro studies were performed to assess the impact of GdFM-Cy3 on cell function and survival. 500,000 GdFM-Cy3-labeled ES-CPCs or control ES-CPCs were injected into the myocardium of mice with and without myocardial infarction. Mice were imaged (9.4T) before and over a two-week time interval after stem cell transplantation. Mice were then euthanized and their hearts were sectioned for fluorescence microscopy. Results In vitro studies demonstrated that GdFM-Cy3 was easily transfectable, non-toxic, stayed within cells after labeling and could be visualized using MRI and fluorescence microscopy. In vivo studies confirmed the efficacy of the agent for the detection of cells transplanted into the hearts of mice after myocardial infarction. A correspondence between MRI and histology was observed. Conclusions The results of the current study suggest that it is possible to identify and potentially track GdFM-Cy3-labeled ES-CPCs in murine infarct models via MRI.

Labeling of cynomolgus monkey bone marrow-derived mesenchymal stem cells for cell tracking by multimodality imaging

“…Mesenchymal stem cells (MSCs) are ethical, practical and biologically appropriate cell populations for cell therapy. In numerous animal and human studies, MSCs show significant potential for tissue regeneration, which requires biosafety and an effective method to detect transplanted cells in vivo [4][5][6][7]. Cell tracking in vivo is an important aspect for the development of successful stem cell therapies.…”

Cardiac cell therapy: current status and future trends

“…These fi ndings support the use of this tool for serial tracking of cell grafts in the myocardium. Techniques to evaluate the timing and specifi c role of SCs labeling with genetic (Tang et al ., 2009;Higuchi et al ., 2009) and immunofl uorescence (Adler et al ., 2009) detectable tags are being investigated in animal models. Lineage fate mapping (Synnergren et al ., 2008;Chiriac et al , 2010) has proved to be an informative tool, and further studies in animal models and ex vivo SC labeling of cells for therapy will continue to be valuable.…”

Computational Analysis: A Bridge to Translational Stroke Treatment

“…To noninvasively monitor implanted NSCs, several labeling techniques have been utilized including, MRI T1-shortening agents (i.e., gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA) labeling) [ 71 ] , positron emitting isotopes in PET [ 72 ] , and MRI T2 shortening contrast agents (i.e., Feridex, iron oxide labeling) [ 73 ] . We have utilized iron oxide labeling of NSCs to noninvasively visualize these cells as they migrate and replicate in ischemic tissue in translational HII models, even as long as 58 weeks after injury [ 2,73 ] .…”