title: Large-scale perfusion culture process for suspended mammalian cells that uses a centrifuge with multiple settling zones.
abstract: A high-cell-density perfusion culture process, using a novel centrifuge, was developed. The centrifuge has spiral multiple settling zones to separate cells from culture medium. Because of the multiple zones, the separation area can be efficiently increased without enlarging the diameter of the centrifuge. The centrifuge used in this study had a separation capacity of 2600 ml culture medium min -1 at 100 g of the centrifugal force. A new cell separation and withdrawal method was also developed. The cells separated in the centrifuge can be withdrawn easily from the centrifuge with no cell clogging by feeding a liquid carrier such as a perfluorocarbon into the centrifuge and pushing the cells out with the liquid carrier. By this culture process, monoclonal antibodies were produced with mouse-human hybridoma X87X at a cell density of about 8 x 10(6) cells ml -1 for 25 days. This centrifuge culture shows promise as a large-scale perfusion culture process. (Himmelfarb et al. 1969; Feder and Tolbert 1983) usually have the problem of filter clogging, and perfusion culture processes with gravitational cell settling (Tokashiki and Arai 1989) are limited in scale-up because of low separation efficiency. Recently centrifugal separation of cells from culture medium has been used for perfusion culture processes (Hamamoto et al. 1989; Tokashiki et al. 1990; Apelman 1992; Jäger 1992). This process essentially has a high separation efficiency and can avoid further operational problems, such as filter clogging, by the type of filtration. To investigate the scale-up feasibility of the centrifuge culture system, we have developed a new type of centrifuge with four layers of spiral cell-settling zones to increase the separation area, and a novel cell separation and withdrawal method using a perfluorocarbon.
authors: Biotechnology Research Laboratories, Teijin Limited, Tokyo, Japan.
authors: H
authors: H
authors: Takamatsu
authors: K
authors: K
authors: Hamamoto
authors: K
authors: K
authors: Ishimaru
authors: S
authors: S
authors: Yokoyama
authors: M
authors: M
authors: Tokashiki
mesh_terms: D000818:Animals; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods; D002478:Cells, Cultured; D002498:Centrifugation / Q000295:instrumentation*; D004867:Equipment Design; D005240:Feasibility Studies; D005465:Fluorocarbon Polymers; D005466:Fluorocarbons; D006801:Humans; D006825:Hybridomas / Q000166:cytology; D008322:Mammals / Q000033:anatomy & histology*; D051379:Mice; D010477:Perfusion / Q000379:methods*
chemical_list: D005465:Fluorocarbon Polymers; D005466:Fluorocarbons; C429307:fluorocarbon 77