Cells-on-chip technologies

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 (chips). These will be applied to a model of the tumor stem cell (CSC) compartment to study their interaction with immune cells (tumor-associated macrophages, TAMs) and elucidate the role of the endocannabinoid system (ES) in the cross-talk between the two populations. Due to their ability to induce aberrant M2 polarization in TAMs, which protects tumor cells from antitumor therapies, CSCs are responsible for the progression and chemoresistance of cancers. Studying the involvement of the ES in the CSC/TAM interaction is of great importance, given that both its immunomodulatory action and its role in the progression and invasiveness of numerous tumors have been documented. To study this interaction, a completely passive microfluidic platform will be developed, in which the careful design of geometric and fluid dynamic parameters will enable the establishment of cell/cell interaction phenomena within it. The chip will feature a central chamber for culturing CSCs in a 3D micromass (tumorsphere) within hydrogel matrices, and lateral chambers for the insertion of TAMs. Microchannel barriers will separate the central chamber from the lateral ones, providing containment of the gel phase, regulating TAM migration toward the gel containing the CSCs, and controlling diffusive transport phenomena. The TAMs, subjected to cytokine-mediated recruitment by the CSCs, will migrate toward the tumor site, invading the hydrogel matrix, similar to what occurs in vivo. The chip will be optimized both to facilitate long-term cell seeding and culture procedures and to enable highly reproducible and precise quantification of morphological and kinetic data of CSC/TAM interaction, down to the single-cell level. To develop a simplified yet standardized model of TAM, a human histiocytic lymphoma myeloid cell line (U937 cells), capable of assuming a macrophage phenotype, will be used. This cell line, engineered to express fluorescence at different wavelengths depending on M1/M2 polarization, will allow for on-chip monitoring of TAM kinetics and polarization modulation with single-cell detail using time-lapse microscopy (TLM). Biological samples collected during and after the experiments will allow for biochemical and molecular biological assessments. The project aims to elucidate the involvement of the ES in the immune response to tumor stem cells, increasing knowledge of a crucial aspect for the outcome of cancer therapies through the development of cell-on-chip technologies capable of mimicking the tumor microenvironment in vitro.

Start and end date

2013-2014

Project Manager

prof Marcella Trombetta, coordinator

Prof. Mauro Maccarrone, consultant to the coordinator

Coordinating institution of the project

Università Campus Bio-Medico di Roma

Other Institutions involved

• National Research Council (CNR)

• University of Rome Tor Vergata

Funding source(s).

Ministry of Education, University and Research (PRIN)