Fig. 1: Development of PDAC-on-a-Chip models.
From: A microfluidic-based PDAC organoid system reveals the impact of hypoxia in response to treatment
A Image of the OrganoPlate® 2-lane from MIMETAS and (B) a zoomed in schematic overview of one chip. The gel inlet (A1) is connected to the gel channel (blue). The perfusion channel (red) connects the perfusion inlet (A2) with the perfusion outlet (A4). C Schematic representation of a chip filled with PDAC organoids and PSCs. The cells are mixed with extracellular matrix, and these are seeded in the gel channel upon pipetting into the gel inlet and subsequently distributed along the gel channel due to capillary forces. The channels are separated by Phaseguides, capillary pressure barriers, which keep the channels separate from each other and allows the stratified loading of culture components. After gelation, cell culture medium was added to the medium inlet and outlet. During all experiments, when the plates were in the incubator (37 °C), these were kept on an (D) interval rocker at an inclination of 7° and an 8-min interval. The interval rocker ensured perfusion of the cultures. To enhance optical clarity, 50 μl of Hanks Balanced Salt Solution (HBSS) (Sigma, 55037 C) was dispensed into the observation windows of the OrganoPlate® 2-lane. E Timeline of the experiment: On day 0 PDAC organoids and PSCs were seeded in Matrigel suspension, cells were allowed to expand until day 4, when these were subjected to chemotherapeutic treatment for 72 h. Cell survival was analyzed using Cell Titer Glo 3D Viability assay. F Representative Phase Contrast (PC) image of a PDAC organoid monoculture in an OrganoPlate 2-Lane. 4x acquisition, Scale bar: 200 um. PC Images were acquired on the ImageXPress Micro XLS Widefield High-Content Analysis System® (Molecular Devices, US).
