The invention relates to an automatic cell culture box. A refrigeration storage device, a CO2 conservation device, a culture medium box or a midway recycling bin, a central incubator, an activation box and a microscope camera structure are respectively connected with a center control system; the refrigeration storage device is arranged on one side in the box body; the CO2 conservation device is arranged on the other side in the box body; the refrigeration storage device and the CO2 conservation device are arranged at an interval by a clapboard in the box body. Compared with the traditional manual culture box, the automatic cell culture box provided by the invention has the advantages that the efficiency is improved by 30 percent, cell culture can be automatically controlled, completely closed culture is realized, an opening step does not exist any more, and the biological safety of cells is thoroughly guaranteed; uniform cell culture standard is realized, cell culture for more people can be simultaneously realized, the efficiency is improved, the labor intensity is reduced, the using amount of consumed materials is reduced, and the culture cost is reduced; the culture box can be used in an ordinary environment, and GMP (Good Manufacturing Practice) building and equipment purchasing costs are greatly reduced.