Study of dendritic cell migration using micro-fabrication

ATP promotes the fast migration of dendritic cells through the activity of pannexin 1 channels and P2X 7 receptors

“…Panx1-dependent signaling is required for the ATP-induced fast migration of DCs Given previous reports suggesting a role for ATP and Panx1 channels in the locomotion of myeloid cells, such as microglia and neutrophils (12,14,33,34), we investigated their possible role in DC motility. For this, we used microfabricated channels that facilitate DC migration by mimicking the confined environment of peripheral tissues and further enable the quantitative analysis of cell speed and cytoskeletal organization (35)(36)(37). We treated DCs with ATP (500 mM) for a short time period (30 min to trigger Panx1 opening), carefully washed the cells, and imaged them while they migrated in the microchannels (see Fig.…”

3D Microfluidic model for evaluating immunotherapy efficacy by tracking dendritic cell behaviour toward tumor cells

“…Both the physical device architecture and the type of 3D matrix, that regulate the phenotype and functions of cells 35, 48 , were taken into consideration for reproducing spaces mimicking in vivo environments where DCs and cancer cells migrate and interact. First, immune-chamber dimensions recreated a spatial confinement, mimicking vessels and tissue interstitial areas, in which DCs, under non-specific adhesive conditions, were able to tune guided migration upon sensing factors released by tumor cells 49 . In addition, the geometry of the connecting-channels allowed DCs to migrate actively through very narrow constrictions to the same extent as when they cross endothelial barriers 50 .…”

“…Therefore, our microfluidic platform was able to recapitulate physical and biochemical environmental cues driving the interactions between DCs and tumor cells, closely resembling in vivo conditions in a time- and spatial-dependent manner. Complementary events, undetectable on 2D flat surfaces used in traditional assays 49 , such as IFN-DC migration and interaction with single cancer cell, were linked in real time using high-resolution time-lapse imaging. Moreover, different parameters characterizing IFN-DC behaviour toward NT and RI-treated cancer cells were evaluated with an automated data analysis system using a revised image analysis algorithm thus overcoming for the first time one of the most important challenges of microfluidic studies, i.e.…”

“…However, the elevated attractant potential of the molecules released by RI-treated dying cells toward IFN-DCs was clearly evident under competition conditions with the massive recruitment of these cells into RI tumor space relative to the NT counterpart. In this case, migrating IFN-DCs adjusted their migratory properties with respect to their functional activity 49 . First, in the immune-chamber they acquired higher pre-channel speed generating crowding in entrance at connecting-channels where they acquired an “amoeboid-like” locomotion mode, with dendritic protrusions at the cell front and contractions at the cell rear, propelling the nucleus through narrow spaces thus allowing persistent cell migration 53 .…”

Developmentally regulated PERK activity renders dendritic cells insensitive to subtilase cytotoxin-induced integrated stress response

“…Microchannels were prepared as previously described (Vargas et al, 2016). For velocity measurements (carried out in 4µm-by-5µm microchannels), phase contrast images of migrating cells were acquired during 16 hours (frame rate of 2 min) on an epifluorescence Nikon Ti-E video microscope equipped with a cooled charge-coupled device (CCD) camera (HQ2, Photometrics) and a 10× objective.…”