Abstract
BackgroundMultiCellular 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.MethodsCell 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.ResultsWe 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.ConclusionOur 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.
Highlights
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
We characterized the cell cycle parameters change in 3D by using pancreatic MCTS models engineered to express cell cycle fluorescent reporters. We report that these MCTS models allow monitoring the activation of cell cycle checkpoints in 3D, providing innovative strategies for the evaluation of new antitumor drugs targeting cell cycle regulation
We demonstrate that classical immunological reagents, as well as engineered cell lines with Fluorescence Ubiquitination cell cycle indicators (Fucci) reporters can be used to precisely describe in "cell cycle words" the complex regionalization that is observed upon spheroid growth in situ
Summary
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. MultiCellular Tumor Spheroid (MCTS) is a 3D model that accurately reproduces the organization of a microtumor, recapitulating cell-cell and cell-microenvironment interactions [1,2]. Spheroids display a gradient of proliferating cells that are located in the outer cell-layers whereas the quiescent cells are located more centrally. This cell heterogeneity is similar to what is found in microregions of a tumor [2,3,4].
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
