Integrated OOC approach to immuno-oncology
The immune system is a striking example of an integrated information system, engaged in coordinated host-protective activities. Organs-on-chip approach (OOC)models allow the direct simultaneous observation of hundreds of different cells, moving, interacting and responding to signals coming from the microenvironment nearby, that give access to a number of parameters describing the system that must be properly measured and elaborated. Combining microfluidics with the ability of cellular imaging enable to collect quantitative data from complex biological systems at a single-cell level.
Reconstitution of the immune-cancer system on chip opens a new window to live observation of the host immune response with or without drug treatments, making OOC a cornerstone for dissecting complex biological phenomena and pre-clinical testing of drugs. Smart implementation of image processing algorithms enable to quantify the simultaneous long-time interactions of huge number of cells and accurately solve the practical problems encountered in multi-cell type context.
Step 1: 2D microfluidic cocultures and observation of cancer–immune cell
motility and interactions
What happens if we put the whole pool of immune cells on chip?
Idea: to monitor and visualize on chip for several days (up to a week) the cellular dynamics ( motility and interactions) of the immune response to melanoma cells in the context of immunocompetence vs immunodeficiency.
Function: support the culture of heterotypic populations (adherent and floating cells) in close proximity to each other, making it suitable to monitor in real-time multicellular events under co-culture conditions.
Design: The PDMS platform consists of two center end-closed channels adjacent to two cell culture compartments (100 µm high) connected via four sets of micron-size grooves (500 x 12x 10 µm). All microfluidic devices were fabricated following well established replica molding procedures.
Loading: Fluorescently labeled B16 F10 melanoma cells and mice spleen cells were added respectively in the left (melanoma compartment) and right chamber (spleen compartment) via the two circular reservoirs (Fig. 1A, green and red arrows). The dead end center channels (containing medium) have been designed to function as buffer chambers (Fig 1B).
Step 2: 3D cancer–immune microenvironments
Step 3: Testing Anti-cancer and Immunotherapy strategies combination on chip
Immunotherapy relies on the use of therapeutic agents that are able to potentiate immune effector mechanisms also inside the tumor microenvironment (TME). DCs have the specific role of recognizing cancer cells, taking up tumor antigens (Ags) and then migrating to lymph nodes for Ag (cross)-presentation to naïve T cells. Interferon-α-conditioned DCs (IFN-DCs) exhibit marked phagocytic activity and the special ability of inducing Ag-specific T-cell response.