The invention discloses an in-vitro 3D tumor microsphere model manufacturing system and method with a controllable core shell structure. The system comprises a mold combination and a biological material perfusion platform; molds for use are directly formed by a 3D printer according to an STL file; the biological material perfusion platform is combined with the mold combination; and a micro pump isused for achieving the accurate perfusion process of materials. The in-vitro 3D tumor microsphere model manufacturing system and method with the controllable core shell structure are based on a 3D printing mold, by means of sequential combination of different molds, by injecting biological materials into different mold cavities, the biological material injected into the molds, cells, medicine orthe above material can be combined freely to form a tumor in-vitro 3D microsphere model with a controllable accurate interior structure and shape characteristic