Abstract: A microfluidic cell culture device is described. The device comprises a microfluidic network comprising a base, a microfluidic channel, and a cover, and at least one perfusion compartment and at least one support compartment inside the microfluidic channel The base and cover each comprise an aperture, thereby defining a conduit through the microfluidic channel. The aperture is in fluidic contact with the at least one support compartment, and with the at least one perfusion compartment through the at least one support compartment. Methods for creating a fluid-fluid interface and for investigating a cellular response to a stimulant using the device are also described.