Patent EP-2639294-A1 (EP)
Inventor: HENON PHILIPPE
IPC: C12M1/00
Filed: 2012-03-15
Abstract: The apparatus comprises tanks of culture medium, growth factors and cells to be cultured, an incubator (12) provided with a thermostated chamber (14) in which a container for cell culture or expansion is housed, a control computer system (24) including a unit for entering and recording data intended to control the culture conditions in the chamber and to manage valves for dispensing fluids in a predefined sequence, and a device (16) for supporting and agitating the cell culture or expansion container that is controlled by the computer system and is housed in the chamber. The apparatus comprises tanks of culture medium, growth factors and cells to be cultured, an incubator (12) provided with a thermostated chamber (14) in which a container for cell culture or expansion is housed, a control computer system (24) including a unit for entering and recording data intended to control the culture conditions in the chamber and to manage valves for dispensing fluids in a predefined sequence, and a device (16) for supporting and agitating the cell culture or expansion container that is controlled by the computer system and is housed in the chamber. The container is formed by a cell expansion bag having an inlet port connected to the tanks and an outlet port connected to a unit (42) for harvesting and storage of the cells after culture. The harvesting and storage unit and the tanks are located outside the chamber and connected to the ports of the cell expansion bag by conduits that form a preassembled module placed in the chamber with the cell expansion bag and pass through a wall of the chamber to feed the cell expansion bag with the culture medium, growth factors and cells to be cultured and to harvest the contents of the cell expansion bag in the harvesting and storage unit while maintaining the chamber closed. The cell expansion bag further comprises a sampling outlet connected to a sampling unit located outside of the chamber by a conduit that passes through the wall of the incubator and is a part of the preassembled module. The incubator includes a cabinet having an opening and a sealed closure door, and a conduits passageway unit that is mounted on a peripheral edge of the opening and includes slightly parallel grooves into which the conduits are engaged, where the grooves are covered by the sealed closure door when in the closed position. The tanks of growth factors and cells to be cultured are formed by bags located above the inlet port of the cell expansion bag so that the content of each of the bags of growth factors and of cells to be cultured flows to the cell expansion bag by gravity. The harvesting and storage unit comprises bags partially located below the outlet port of the cell expansion bag so that the content of the cell expansion bag flows to the bags of the harvesting and storage unit by gravity after culturing. The cell expansion bag comprises flexible liquid-tight walls that are permeable with gas such as carbon dioxide. The walls of the cell expansion bag have properties minimizing the adhesion of the cells to be cultured to the walls. The apparatus further comprises a peristaltic pump for controlling the supply of the culture medium to the cell expansion bag and to the tanks of growth factors and of cells to be cultured and for rinsing the tanks, and two bags forming air trap, where one of the bags is connected to the tanks of growth factors and of cells to be cultured, the other bag is connected to the cell expansion bag, and the two bags collect and store the air contained in the conduits, the cell expansion bag and/or the tanks. The conduits are formed by flexible tubes, where some of the tubes pass through valves intended to pinch the tubes in the closed position. The supporting and agitation device comprises a tray to support the cell expansion bag that is mounted in rotation around a first horizontal axis and is movable around the axis between a slightly horizontal position for cell culture and a slightly vertical position for harvesting the cells after culture. The tray is mounted in rotation around a second horizontal axis around which the tray is intended to oscillate for agitation and homogenization of the content of the cell expansion bag. The tray bears valves for controlling the supply of the cell expansion bag, the harvesting of the content of the cell expansion bag, and the sampling of the cell expansion bag. The supporting and agitation device comprises a vertical arm having at its upper end unit for attaching the bag forming an air trap connected to the cell expansion bag. The harvesting and storage unit is mounted in rotation around a horizontal axis and movable around the axis between a slightly vertical position and a slightly horizontal position in which the harvesting and storage unit is located completely below the cell expansion bag. Independent claims are included for: (1) a sterile and disposable cell culture kit; and (2) an automated method for cell culture.