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Open Access Highly Accessed Research article

Multicellular tumor spheroid models to explore cell cycle checkpoints in 3D

Jennifer Laurent12, Céline Frongia12, Martine Cazales12, Odile Mondesert12, Bernard Ducommun123 and Valérie Lobjois124*

Author Affiliations

1 Université de Toulouse; ITAV-USR3505, F-31106, Toulouse, France

2 CNRS; ITAV-USR3505, F-31106, Toulouse, France

3 CHU de Toulouse, F-31059, Toulouse, France

4 ITAV - USR3505, Centre Pierre Potier, 1 place Pierre Potier, 50624, 31106, Toulouse Cedex 1, BP, France

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BMC Cancer 2013, 13:73  doi:10.1186/1471-2407-13-73

Published: 8 February 2013

Abstract

Background

MultiCellular Tumor Spheroid (MCTS) mimics the organization of a tumor and is considered as an invaluable model to study cancer cell biology and to evaluate new antiproliferative drugs. Here we report how the characteristics of MCTS in association with new technological developments can be used to explore the regionalization and the activation of cell cycle checkpoints in 3D.

Methods

Cell cycle and proliferation parameters were investigated in Capan-2 spheroids by immunofluorescence staining, EdU incorporation and using cells engineered to express Fucci-red and -green reporters.

Results

We describe in details the changes in proliferation and cell cycle parameters during spheroid growth and regionalization. We report the kinetics and regionalized aspects of cell cycle arrest in response to checkpoint activation induced by EGF starvation, lovastatin treatment and etoposide-induced DNA damage.

Conclusion

Our data present the power and the limitation of spheroids made of genetically modified cells to explore cell cycle checkpoints. This study paves the way for the investigation of molecular aspects and dynamic studies of the response to novel antiproliferative agents in 3D models.

Keywords:
Spheroid; 3D; Cell cycle; Checkpoint; Fucci reporters