Figure 5 – CRC eSF microfluidic model. A) Schematics of the CRC microfluidic model containing endothelial cells seeded on the microchannel and colorectal cancer cells encapsulated throughout the eSF hydrogel. B) Brightfield images of HCoMECs perfused and adhered in the microchannel on Day 1 and Day 3; and Fluorescence microscopy images of HCoMECs after 5 days of culture (DAPI, blue, nuclei; phalloidin, F-actin, red) showing endothelial cells aligning and creating an endothelialized lumen-like structure within the microchannel. C) 3D confocal image of a Live/dead at Day 1. D) Live/dead assay on days 1 and 3. Live cells are stained in green and dead cells in red. Scale bar: 100 µm. E) Cell viability studies. ATP quantification graph showing HCT-116 cell metabolic activity in 2D and 3D (encapsulated in eSF hydrogels) cultures after 1, 3 and 7 days of culture. Data are presented as mean ± stdev (n = 3), (*) denotes statistical differences in the same day in different conditions (p < 0.05), (#) denotes statistical differences within the same state, but compared to timepoint day 1. F) Alamar blue performed in 14% eSF microfluidic platforms with HCT-116 cells encapsulated within, with an 8 µL per hour flow rate of Gemcitabine solution at days 1, 3 and 7. * significant differences were observed when comparing the control to Gemcitabine (GEM) condition within the same timpoint. # significant differences when comparing the same situation to the previous timepoint. p<0.05.