Design and construction of the parallel-plate coculture flow chamber.
(A) Components of the chamber. Two sets of glass slides (1 and 5) and silicone gaskets (2 and 4) and the polycarbonate insert holder (3) are held together by a vacuum suction applied at the perimeter of the side via ports v and vi, forming a channel inside the lower silicone gasket (4). The insert holder is machined precisely to allow the introduction of the EC/SMC coculture module (viii) into the channel. Cultured medium enters at port i through slit iii into the channel, and exits through slit iv and port ii. The chamber allows on-line microscopic visualization of the cells during the flow experiment via the observation window (vii) opened in the upper silicone gasket (2). (B) Schematic diagram of the parallel-plate coculture flow channel. ECs were seeded onto the inverted side of the membrane containing 0.4-μm pores configured at a density of 1.6 × 106 pores/cm2 and then subjected to flow, while SMCs cultured in the opposite side of the membrane were maintained in a static condition.
