The invention relates to a stem cell culture device, and belongs to the technical field of stem cell culture. The stem cell culture device includes a box body, the box body is provided with a liquid storage tank, a liquid pump and a liquid outlet pipe, the liquid storage box is connected with the liquid pump, the liquid pump is provided with a liquid outlet pipe, the liquid outlet pipe is providedwith a horizontal pipe, the horizontal pipe is provided with a spray head, the box body is provided with a rotating motor, the rotating motor is provided with a turntable, the turntable is provided with an insert hole, the insert hole is matched with the spray head, the insert hole is provided with a limiting structure, the box body is provided with an air inlet pipe and an air outlet pipe, the air inlet pipe is provided with a casing pipe, the casing pipe is provided with a movable plate, the casing pipe and the movable plate are provided with through holes, the casing pipe is provided withan adsorbent, the air inlet pipe is provided with a mounting frame, the mounting frame is provided with an induced draft fan, the air outlet pipe is provided with a one-way control structure, the boxbody is provided with a door panel, the door panel is provided with a sealing gasket for sealing, and a glass window for observation is fixedly installed in a through groove of the door panel. A culture dish is inserted into the insert hole, a second spring of the limiting structure pushes a movable block to abut against the culture dish so as to limit the culture dish in the insert hole, the stability of the culture dish is ensured, and the stability of stem cell culture is ensured.