Cells-on-chip technologies

Cells-on-chip technologies for the study of the endocannabinoid system in an in vitro model of tumor/immune system interaction

Project Objectives

The project aims at developing advanced models for the in vitro study of cellular interactions by exploiting 3D co-culture technologies within microfluidic devices (chips). These models will be applied to an experimental model of the cancer stem cell (CSC) compartment, in order to study the interaction of the CSC with immune system cells (Tumor-Associated Macrophages, TAM) and elucidate the role of the endocannabinoid system (SE) in the cross-talk between these two cell populations.  

Cancer stem cells are responsible for progression and chemoresistance in neoplasia due to their ability to cause aberrant and protective polarization of cancer cells against anticancer therapies in TAM the M2. The study of the involvement of SE in CSC/TAM interplay has high importance due to its well-known immunomodulatory action as well as to its role in the progression and invasiveness of several types of tumors. 

To study this involvement, an entirely passive microfluidic platform will be developed, where careful design of the geometrical and fluid dynamic parameters will allow the establishment of cell/cell interaction phenomena inside this platform. The chip will have a central chamber for a 3D micromass culture of CSC (tumorsphere), within the hydrogel matrices, and side chambers for loading TAM. The central chamber will be separated from the side chambers through microchannel barriers so generating the containment of the gel phase, the regulation of TAM migration to the CSC containing gel and control over diffusive transport phenomena. TAM, through the cytokine-mediated attraction of CSC, will migrate to the tumor site so invading the hydrogel matrix, which is similar to the in vivo process. 

The chip will be optimized to facilitate the procedures of long-term cell plating and culturing, as well as to enable a high reproducibility and accurate quantification of morphological and kinetic data on CSC/ TAM interplay at single cell level. To obtain a simplified and standardized TAM model, a myeloid cell line of human histiocytic lymphoma (U937 cells), which is a model able to acquire a macrophage phenotype, will be used. This cell line, modified to express fluorescence signals with different wavelength depending on the M1/M2 polarization, will allow the study to follow kinetics and modulation of TAM polarization on-chip at a single cell level by using time-lapse microscopy (TLM). 

Biological sampling will be performed during and after the experiments to enable biochemical and molecular-biological evaluations. The project aims to elucidate the involvement of SE in the immune response to cancer stem cells so increasing our knowledge of a fundamental feature for the outcome of cancer treatments, by virtue of the development of cells-on-chip technologies able to mimic tumor microenvironment in vitro.

Start/End date

2013 - 2014

Principal Investigators

  • Prof. Marcella Trombetta - Coordinator
  • Prof. Mauro Maccarrone - Consultant to the coordinator

Host Institution

Campus Bio-Medico University of Rome

Other Institutions involved

  • Università di Roma “Tor Vergata”.
  • Consiglio Nazionale delle Ricerche (CNR) [National Research Council (NRC)]

Source of funding

Italian Ministry for Education