It is an object to achieve a closed system that has a simple structure without resorting to large scale equipment and that sequentially carries out all of the step of separating useful cells, the step of culturing the useful cells separated in the separation step, and the step of washing and concentrating the cells cultured in the culturing step and to enable the preparation of useful cells having high safety and quality, while improving operability. A disposable set for cell culture includes a cell culture container (CC) having a liquid inlet (Lin) and a liquid outlet (Lout), a cell separation kit (A) that is for separating cells used for cell culture and is connected to the liquid inlet (Lin), and a cell collection kit (B) that is for washing and concentrating cultured cells in the cell culture container (CC) and is connected to the liquid outlet (Lout), and a cell culture device uses the disposable set for cell culture and includes flow path on-off valves and pumps provided at appropriate positions of pipelines of the disposable set and a controller controlling them.