Cell culture system with manifold

A cell culture apparatus having a cascading manifold, comprising:

• a plurality of cell culture chambers, each cell culture chamber comprising a top surface and a bottom surface and configured to culture cells on the bottom surface, each cell culture chamber comprising gas permeable, liquid impermeable film;
• wherein the plurality of cell culture chambers are stacked one on top of the other to form a stacked cell culture unit, each cell culture unit comprising a manifold;
• a plurality of cell culture units stacked one on top of the other;
• each manifold comprising an inlet, an overflow outlet and a plurality of cell culture chamber openings in fluid communication with each of the plurality of cell culture chambers of a cell culture unit;
• wherein fluid, introduced into a first manifold through the inlet, flows into each cell culture chamber through the plurality of cell culture chamber openings until each cell culture chamber is filled with fluid and then, once each cell culture chamber is filled, the first manifold fills with fluid and fluid flows out of the first manifold through the overflow outlet;
• wherein the overflow outlet of the first manifold of a first cell culture unit is coupled to the inlet of a second manifold of a second cell culture unit;
• wherein fluid flowing out of the first manifold through the overflow outlet flows into the inlet of a second manifold of a second cell culture unit.