Abstract
BackgroundWe investigated the utility of bioluminescence imaging (BLI) using firefly luciferase in monoclonal and polyclonal populations of leukemia cells in vitro and in vivo.MethodsMonoclonal and polyclonal human lymphoid and myeloid leukemia cell lines transduced with firefly luciferase were used for BLI.ResultsKinetics and dynamics of bioluminescence signal were cell line dependent. Luciferase expression decreased significantly over time in polyclonal leukemia cells in vitro. Transplantation of polyclonal luciferase-tagged cells in mice resulted in inconsistent signal intensity. After selection of monoclonal cell populations, luciferase activity was stable, equal kinetic and dynamic of bioluminescence intensity and strong correlation between cell number and light emission in vitro were observed. We obtained an equal development of leukemia burden detected by luciferase activity in NOD-scid-gamma mice after transplantation of monoclonal populations.ConclusionThe use of monoclonal leukemia cells selected for stable and equal luciferase activity is recommended for experiments in vitro and xenograft mouse models. The findings are highly significant for bioluminescence imaging focused on pre-clinical drug development.
Highlights
We investigated the utility of bioluminescence imaging (BLI) using firefly luciferase in monoclonal and polyclonal populations of leukemia cells in vitro and in vivo
Instability and incomparability of luciferase activity in polyclonal human leukemia cell lines in vitro Polyclonal luciferase expressing populations of human T-cell acute lymphoblastic leukemia (Jurkat), B-cell acute lymphoblastic leukemia (697) and chronic myeloid leukemia (K562) cell lines were generated and luciferase activity was determined by measurement of bioluminescence intensity
We have shown that the bioluminescence signal intensity and the dynamics of luciferase activity in vitro were cell line dependent (Figure 1)
Summary
We investigated the utility of bioluminescence imaging (BLI) using firefly luciferase in monoclonal and polyclonal populations of leukemia cells in vitro and in vivo. Further mammalian tissue is known to be a turbid medium that both scatters and absorbs photons This is mostly due to changes in refractive index at cell membranes and internal organelles, and can lead to a scattered and attenuated bioluminescence signal, which has influence on investigations especially in deeper tissue [18]. Bioluminescence imaging using firefly luciferase in vitro and in vivo is often performed with potentially unstable luciferase-expressing polyclonal cell populations. In this study we investigated the limitations, advantages and disadvantages of bioluminescence imaging using a firefly luciferase system with monoclonal and polyclonal human leukemia cell populations in vitro and in a xenograft mouse model
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