Cells-on-chip technologies for the study of the endocannabinoid system

Project objectives

The project aims to develop advanced models for the in vitro study of cellular interactions, leveraging 3D co-culture technologies within microfluidic devices. Specifically, on-chip technologies are being applied to a tumor stem cell (CSC) model to study their interaction with immune cells (Tumor-Associated Macrophages, TAMs) to elucidate the role of the endocannabinoid system (ES) in the cross-talk between the two populations. CSCs are responsible for the progression and chemoresistance of cancers due to their ability to induce a shift in TAM polarization state (from M1 to M2), which no longer perform their cytotoxic function but instead protect tumor cells against antiblastic therapies. Studying the involvement of ES in CSC/TAM interaction is of great importance, given that both their immunomodulatory action and their role in the progression and invasiveness of numerous tumors have been documented.

The on-chip technology developed in this project serves as a platform for single-cell studies of CSC/TAM interaction. The chips feature a central chamber for culturing CSCs in a 3D micromass (tumorsphere) within hydrogel matrices; lateral channels allow the insertion of TAMs, which, under cytokine-mediated stimulation, migrate toward the tumor site, invading the hydrogel matrix, similar to what occurs in vivo. The chip design is optimized to facilitate long-term cell seeding and culture procedures, while also enabling highly reproducible and precise quantification of morphological and kinetic data of CSC/TAM interaction.

To obtain a simplified yet standardizable model of TAM, a human histiocytic lymphoma myeloid cell line (U937 cells), capable of assuming a macrophage phenotype, is used. It has been engineered to express fluorescence at different wavelengths depending on M1/M2 polarization, allowing the kinetics and polarization modulation of TAMs to be followed on-chip. Experiments are conducted using time-lapse microscopy (TLM), monitoring migration and polarization phenomena at the single-cell level. The on-chip approach allows both the collection of microvolumes of culture supernatants and the recovery of viable cells for subsequent biochemical and molecular biological characterization of the HS components. The project aims to elucidate the involvement of the ES in the immune response to tumor stem cells, increasing knowledge on an aspect of fundamental importance for the outcome of cancer therapies, by virtue of the development of cell-on-chip technologies capable of mimicking, in vitro, the tumor microenvironment.  

Start and end date

March 2014 - March 2017

Project Manager

prof Marcella Trombetta - Project Manager

Coordinating institution of the project

Università Campus Bio-Medico di Roma

Other Institutions involved

• CNR-IFN Institute of Photonics and Nanotechnologies

• UTV Unit - Biochemistry Lab, Department of Experimental Medicine and Surgery, University of Rome "Tor Vergata"

Funding source(s).

PRIN 2012