Different configurations for studying paracrine and physical interactions between two different cell populations (green and orange). Conventional macroscale approaches without intercellular distance control: seeding in (a) 2D and (c) 3D. Micropatterning approaches with intercellular distance control: (b) 2D and (d) 3D. Microfluidic cell culture of different cell populations: (e) in 3D scaffolds under microenvironmental control and (f) for tissue specimen culture or forming organized cellular structures.
Microfluidic device for culturing mural cells and endothelial cells. (a) Coculture model including mural cells and EC (HMVEC). (b) SMCs are cultured in the right channel and ECs are cultured in the center channel. The presence of ECs increased 3D invasion of SMCs into the collagen type I ECM, while ECs become stabilized by the presence of SMC (Ref. 21). (c) Adhered SMCs on the HMVEC monolayer, shown by a white arrowhead. Nucleus (DAPI; blue) and actin filaments (rhodamine phalloidin; yellow). Green antibody staining indicates HMVECs.
Microfluidic device for integrating different liver cells. (a) Coculture model including hepatocytes and EC. Hepatocytes are cultured in the top channel in the figure and form 3D structures adjacent to the collagen type I gel between the top and the bottom channels. ECs are cultured in the bottom channel, and they migrate into the gel to form capillary networks. (b) Triculture model including hepatocytes, HSC, and EC. HSCs are introduced before adding ECs. Phase contrast images show that three cell types behave differently in the different regions 1–3.
Microfluidic device for recreating TC-EC interactions. (a) Schematic of the coculture model. (b) Brain cancer cells (U87MG) are cultured in the right channel and microvascular dermal endothelial cells are cultured in the center channel. U87MG invade into the collagen type I gel and approach the endothelial cells. (c) Protruding U87MG toward the endothelial monolayer. [(d) and (e)] Time-lapse images (fluorescent and phase-contrast) every 24 h of U87MG transmigrating into EC channel. Transmigrated cell marked with blue arrowhead. Nucleus (DAPI; blue) and actin filaments (rhodamine phalloidin; yellow). Green staining indicates GFP expressing brain cancer cells.
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