Research on mass culture of cell from the viewpoint of chemical engineering

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1. High density culture of anchorage-dependent mammalian cell using polyurethane foam as a substratum for cell attachment and production of useful biologicals Anchorage-dependent mammalian cells were cultivated at high cell density in a novel culture system using polyurethane foam (PUF) as a substratum for cell attachment. PUF has a macroporous structure giving a high surface area to volume ratio. Monkey kidney cells (Vero) attached to the internal surface of PUF and grew to a high cell density (1.04x10^8 cells/cm^3 PUF) in PUF stationary cultures. Furthermore, Vero cells were found to produce urokinase stored in the cell. In PUF stationary culture, Vero cells produced the urokinase from the early logarithmic phase of cell growth. The specific activity of urokinase became highest in the late-logarithmic phase and attained to 70 I.U./mg-protein. This maximal specific activity was about three times as large as that in petri dish culture. These results suggested that the urokinase product … More ivity of Vero cells increased by high density cell culture using PUF.2. Liquid spouted bed culture of microencapsulated mammalian cells Microencapsulated MPC11 cells, which are anchorage-independent, were tried to propagate in a newly designed liquid spouted bed culture system. As the culture system was suitable for separation of medium and microcapsules, the medium could be circulated sufficiently to give nutrients, oxygen, etc. to microencapsulated cells and wash out waste. As a result, the maximum cell density in the microcapsules reached comparatively high 3.3x10^7 cells/ml.3. Production and purification of carotenoid pigments from high density culture of carrot cells A method for purification of carotenoid pigments from cultured carrot cells was established. The relation between cell growth and carotenoid production of carrot cells in shake culture and air-lift culture was studied by using the purification method. The results indicated that carotenoids were growth-dependently produced from the early logarithmic phase of cell growth, and that productivity decreased in the stationary phase, even if medium was exchanged in the sationary phase. Finally, 29.4 mu g/liter-medium of beta-carotene was obtained from high density cultured carrot cells using an air-lift column of 1 liter volume equipped with a draft tube. Less