Dynamic breeding, proliferation and/or differentiation of suspended primary or stem cells of human and animal origins, comprises adhering cells on cell carrier in rotating bed-bioreactor and breading by alternating movement of cell carrier

摘要

Dynamic breeding and/or proliferation and/or differentiation of suspended primary cells or stem cells of human and animal origins in a rotating bed bioreactor, which has a cell carrier (5) formed as rotating beds and works under the conditions of e.g. glycerol 3-phosphate (GLP) and guanidine condensed phosphate (GCP), comprises adhering the primary cells or stem cells suspended in a culture medium on the cell carrier in a rotating bed-bioreactor and then dynamically breading and expanding by alternating movement of the cell carrier by the culture medium and by the overlay-atmosphere. Dynamic breeding and/or proliferation and/or differentiation of suspended primary cells or stem cells of human and animal origins in a rotating bed bioreactor, which has a cell carrier (5) formed as rotating beds and works under the conditions of glycerol 3-phosphate (GLP), guanidine condensed phosphate (GCP), guanosine monophosphate (GMP) and guanosine triphosphate (GTP), comprises adhering the primary cells or stem cells suspended in a culture medium on the cell carrier in a rotating bed-bioreactor and then dynamically breading and expanding by alternating movement of the cell carrier by the culture medium and by the overlay-atmosphere. The adhering of stem cells or primary cells takes place in a closed system of a rotating bed bioreactor without the movement of the rotating bed and without the addition of fresh culture medium over a period of 1 to 12 hours, preferably 3 to 6 hours, one at a time on both sides of the cell carrier. A small volume of culture medium located outside of the rotating bed is pumped in such a manner that it is not overflowed with culture medium, but fills the head space of the rotating bed bioreactor, preferably 5-10% of the volume of the reactor vessel (1) with a gas mixture of carbon dioxide and air and/or nitrogen and the pH and oxygen pressure value is set in dependence to the proliferating cells, preferably the oxygen pressure value is set to 1-100%. An independent claim is also included for a device for carrying out the above method, comprising a reactor vessel with at least one removable cap, controllable port A and B for the inflow and outflow of medium and ports C and D for the supply and removal of overlay-atmosphere and a contact-driven rotating shaft (7) axially arranged in the reactor vessel, over a magnetic drive, where on the shaft, at least on cell carrier is arranged, which is moved by the contact-less magnetic drive alternating by a culture medium and by the overlay atmosphere located in the head-space of the reactor vessel. The rotatable shaft in the reactor vessel is placed in a removable manner on one side and stored in a cover on the other side, which is removable from the reactor vessel. The rotating bed consists of a packet of cell carrier of rectangular discs, which are arranged over one another, with a defined distance from each other and parallel to the shaft of the rotating bed and have different widths, which match respectively with the diameter of the reactor vessel and has a thickness of 0.2-2 mm, preferably 0.5-1 mm, and circular end plates with appropriately sized grooves to fix the rectangular discs, where these are placed in a defined distance of 0.2-4 mm, preferably 0.4-0.8 mm, where the distance between the cell carrier ensures accurate by appropriate knobs on the disc surfaces.