A perfusion cell culture device includes a driving module and a plurality of cell culture modules. The driving module includes a driving source connecting opening and a chamber. The chamber and the driving source connecting opening are connected. Each of the culture modules includes a fluid channel, a first elastic element, two flow direction controlling units and a cell culture zone. The fluid channel is disposed above the chamber, the first elastic element is disposed between the fluid channel and the chamber, the two flow direction controlling units are respectively disposed on two ends of the fluid channel and connected to the fluid channel selectively, and the cell culture zone is connected to the two flow direction controlling units. |