Imaging cellular pharmacokinetics of (18)F-FDG and 6-NBDG uptake by inflammatory and stem cells

OBJECTIVES: Myocardial infarction (MI) causes significant loss of cardiomyocytes, myocardial tissue damage, and impairment of myocardial function. The inability of cardiomyocytes to proliferate prevents the heart from self-regeneration. The treatment for advanced heart failure following an MI is heart transplantation despite the limited availability of the organs. Thus, stem-cell-based cardiac therapies could ultimately prevent heart failure by repairing injured myocardium that reverses cardiomyocyte loss. However, stem-cell-based therapies lack understanding of the mechanisms behind a successful therapy, including difficulty tracking stem cells to provide information on cell migration, proliferation and differentiation. In this study, we have investigated the interaction between different types of stem and inflammatory cells and cell-targeted imaging molecules, (18)F-FDG and 6-NBDG, to identify uptake patterns and pharmacokinetics in vitro. METHODS: Macrophages (both M1 and M2), human induced pluripotent stem cells (hiPSCs), and human amniotic mesenchymal stem cells (hAMSCs) were incubated with either (18)F-FDG or 6-NBDG. Excess radiotracer and fluorescence were removed and a 100 μm-thin CdWO(4) scintillator plate was placed on top of the cells for radioluminescence microscopy imaging of (18)F-FDG uptake, while no scintillator was needed for fluorescence imaging of 6-NBDG uptake. Light produced following beta decay was imaged with a highly sensitive inverted microscope (LV200, Olympus) and an Electron Multiplying Charge-Couple Device (EM-CCD) camera. Custom-written software was developed in MATLAB for image processing. RESULTS: The average cellular activity of (18)F-FDG in a single cell of hAMSCs (0.670±0.028 fCi/μm(2), P = 0.001) was 20% and 36% higher compared to uptake in hiPSCs (0.540±0.026 fCi/μm(2), P = 0.003) and macrophages (0.430±0.023 fCi/μm(2), P = 0.002), respectively. hAMSCs exhibited the slowest influx (0.210 min(-1)) but the fastest efflux (0.327 min(-1)) rate compared to the other tested cell lines for (18)F-FDG. This cell line also has the highest phosphorylation but exhibited the lowest rate of de-phosphorylation. The uptake pattern for 6-NBDG was very different in these three cell lines. The average cellular activity of 6-NBDG in a single cell of macrophages (0.570±0.230 fM/μm(2), P = 0.004) was 38% and 14% higher compared to hiPSCs (0.350±0.160 fM/μm(2), P = 0.001) and hAMSCs (0.490±0.028 fM/μm(2), P = 0.006), respectively. The influx (0.276 min(-1)), efflux (0.612 min(-1)), phosphorylation (0.269 min(-1)), and de-phosphorylation (0.049 min(-1)) rates were also highest for macrophages compared to the other two tested cell lines. CONCLUSION: hAMSCs were found to be 2–3× more sensitive to (18)F-FDG molecule compared to hiPSCs/macrophages. However, macrophages exhibited the most sensitivity towards 6-NBDG. Based on this result, hAMSCs targeted with (18)F-FDG could be more suitable for understanding the mechanisms behind successful therapy for treating MI patients by gathering information on cell migration, proliferation and differentiation.