Multicellular Tumor Spheroids as a Model System for the Evaluation of PET Radiotracer Uptake

Aim:

Three-dimensional multicellular tumor spheroids (MCTS) of tumor cell lines are a promising model system which reflects the in vivo situation of tumor microregions with increasing accuracy compared with conventional monolayer culture. In this study we quantified the uptake of radiotracers [¹⁸F]FDG and [¹⁸F]FMISO in various types of MCTS at different diameters and after defined times in culture.

Material and Methods:

Human tumor cell lines FaDu, HT-29, MCF-7 and EJ-28 and the murine line B16-F10 were cultured in 96 well plates. Culturing was performed in the presence of 0.24% methylcellulose for 3 to 7 days and with seeding densities of 1,000, 5,000, 10,000 and 30,000 cells per well. Diameters of individual spheroids were assessed and cellular uptake of [¹⁸F]FDG and [¹⁸F]FMISO was recorded after an incubation interval of 2 to 4 hours. MCTS were then harvested, washed onto filters and the individual uptake per spheroid was measured with a gamma counter.

Results:

Between day 3 and day 7, spheroids of a starting concentration of 10,000 cells showed an increase in diameter of 1.4-fold for HT-29, 1.2-fold for FaDu, 1.3-fold for MCF-7 and 1.1-fold for both EJ-28 and B16-F10 cells. In all cell lines, we observed a good correlation between uptake of [¹⁸F]FDG and spheroid diameter and protein content, respectively. With an experimental setting of 10.000 initiated cells and 3 days of culturing, the highest [¹⁸F]FDG uptake was achieved in B16-F10 cell spheroids with a value of 1.46±0.41 percent injected dose per MCTS (%ID/MCTS). In comparison, FaDu uptake was 0.63±0.15%ID/MCTS and 0.38±0.05%ID/MCTS in HT-29. EJ-28 and MCF-7 exhibited less than 0.2%ID/MCTS. In parallel, a different uptake pattern was observed for [¹⁸F]FMISO. Highest uptake was documented in B16-F10 with 0.23±0.11%ID/MCTS, whereas all other cell lines showed an uptake of 0.03%ID/MCTS or less. These data indicate larger areas of hypoxia in the cores of B16-F10 spheroids compared to other cell lines.

Conclusion:

Our data show a spheroid-type specific increase in the uptake of both [18F]FDG and the hypoxia marker [¹⁸F]FMISO as a function of the spheroid diameters which is indicative for an increasing amount of metabolically active cells and an increment of hypoxic areas in growing spheroids. We conclude (i) that radiotracer uptake is a promising approach to monitor the pathophysiological situation in MCTS and (ii) that MCTS are a valuable tool to study the uptake of new radiotracers in a pathophysiological tumor milieu.