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Live cell division dynamics monitoring in 3D large spheroid tumor models using light sheet microscopy
Authors:Corinne Lorenzo  Céline Frongia  Raphaël Jorand  Jérôme Fehrenbach  Pierre Weiss  Amina Maandhui  Guillaume Gay  Bernard Ducommun  Valérie Lobjois
Affiliation:1. Université de Toulouse; ITAV-UMS3039, F-31106, Toulouse, France
2. CNRS; ITAV-UMS3039, F-31106, Toulouse, France
3. Université de Toulouse; IMT-UMR5219, F-31062, Toulouse, France
4. CNRS; IMT-UMR5219, F-31062, Toulouse, France
5. CHU de Toulouse, F-31059, Toulouse, France
Abstract:

Background

Multicellular tumor spheroids are models of increasing interest for cancer and cell biology studies. They allow considering cellular interactions in exploring cell cycle and cell division mechanisms. However, 3D imaging of cell division in living spheroids is technically challenging and has never been reported.

Results

Here, we report a major breakthrough based on the engineering of multicellular tumor spheroids expressing an histone H2B fluorescent nuclear reporter protein, and specifically designed sample holders to monitor live cell division dynamics in 3D large spheroids using an home-made selective-plane illumination microscope.

Conclusions

As illustrated using the antimitotic drug, paclitaxel, this technological advance paves the way for studies of the dynamics of cell divion processes in 3D and more generally for the investigation of tumor cell population biology in integrated system as the spheroid model.
Keywords:
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