Microfluidic devices, systems, and methods for investigating three-dimensional cellular structures

A microfluidic device for investigating three-dimensional (3D) cell structures, the microfluidic device comprising:

• a body having a first end, a second end, two lateral edges, an upper side, a lower side, a first layer region between a medial plane of the body and the lower side, and a second layer region between the medial plane and the upper side;
• where the first layer region defines one or more rows of wells, each well including an open top portion at the medial plane and a bottom between the medial plane and the lower side, and where each well has a minimum transverse dimension of 1300 microns;
• where the second layer region defines one or more elongated channels that each include an open bottom portion at the medial plane and a top between the medial plane and the upper side, each elongated channel of the one or more elongated channels aligned with a respective one of the rows of wells such that the open bottom portion of each elongated channel opposes the open top portion of each well in the respective one of the rows of wells such that each elongated channel is in fluid communication with each well in the respective one of the rows of wells;
• and
• where the second layer region further defines a first port, a second port, and a third port that each extend from an exterior surface of the body and are each in fluid communication with an elongated channel of the one or more elongated channels such that a first flow path from the first port to the second port corresponds to a first set of wells of a row of wells aligned with the elongated channel and a second flow path from the second port to the third port corresponds to a second set of wells of the row of wells that is different than the first set of wells.