Patent EP-2543719-A1 (EP)
Title: Meander bioreactor and method for dynamic expansion, differentiation and harvest of hematopoietic cells
Inventor: HOFFMEISTER HANS PROF DR
IPC: C12M3/00
Filed: 2012-07-06
Abstract: Meander bioreactor for dynamic expansion, differentiation, and harvesting of non adherently growing cells, preferably mixtures or isolated fractions of hematopoietic cells, stem cells and progenitor cells of human origin, comprises a rectangular bioreactor vessel (1) made of clear polymeric material, which is closed with a lid (2) in a sterile manner. Inlets (3) and outlets (4) are arranged on the lid for the flow through of medium and gases. The bottom surface of the bioreactor vessel is 20-500 cm 2>, preferably 50-100 cm 2>, and is provided with strip-shaped partitioning walls (11). Meander bioreactor for dynamic expansion, differentiation, and harvesting of non adherently growing cells, preferably mixtures or isolated fractions of hematopoietic cells, stem cells and progenitor cells of human origin, comprises a rectangular bioreactor vessel (1) made of clear polymeric material, which is closed with a lid (2) in a sterile manner. Inlets (3) and outlets (4) are arranged on the lid for the flow through of medium and gases, which are mounted at different heights. The bottom surface of the bioreactor vessel is 20-500 cm 2>, preferably 50-100 cm 2>, and is provided with strip-shaped partitioning walls (11) partitioning the bottom surface and enforcing a meander-like flow through of the bioreactor vessel with gases and medium. The distance of the partitioning walls from one another and the side- and front walls of the bioreactor vessel is selected so that channels formed in the flow filaments by the partition walls forms an average laminar upper flow with a Froude number of less than 0.005, preferably 0.002 and a bottom flow with a Froude number of 0 or close to 0. An independent claim is also included for dynamic differentiating and harvesting of hematopoietic cells, preferably natural killer cells, T-lymphocytes and B-lymphocytes, comprising (a) supplying an amount overlying the inlet and the outlet for the media circulation in medium suspended cells via the inlet to the horizontally erected bioreactor vessel, in a cell count of 0.1-2x 10 6>, preferably 0.5-1x 10 6>cells/cm 2>forcing a complete covering of the bottom surface of the bioreactor vessel and a cell layer with a close contact of the cells with each other, (b) storing the bioreactor vessel for 30-180 minutes, preferably 60-120 minutes in a horizontal position and at rest, (c) switching on the gas supply via the inlet and outlet with a gas rate controlling a laminar and without turbulence generated flow, (d) pumping cell-free culture medium with rates of 1-20 x/hour, preferably 5-15 x/hour, and (e) collected samples periodically via the sterile entry, where fresh medium is supplied via the inlet with continuously increasing amount upon decreasing the glucose content to one third of the initial value in the culture medium so that the glucose content of is greater than the one third of the initial value and the lactate concentration is less than 3 g/liter. The media quantity in the bioreactor does not increase above a factor 3 relative to the amount of the cell suspension at the start of cultivation, and the ratio of the circulating medium quantity does not exceed to the supplied quantity of fresh medium of 5-1.