Pilot production of u-PA with porous microcarrier cell culture.
A recombinant DNA CHO cell line secretingurokinase-type plasminogen activator (u-PA) wascultivated with Cytopore cellulose porousmicrocarriers in a 30l Biostat UC stirred tankreactor. After 26 days of culture, using a spinfilter toretain cells in bioreactor, the cell density couldreach 1.33 x 10(7) ml(-1). The maximal u-PAactivity in supernatant was 7335 IU.ml(-1), and204l supernatant containing 7.1 g u-PA was harvested.After 100 days of culture with 0.1% fetal bovineserum medium, a modified cell retention system whichcan be washed-out backward, substituted thespinfilter to prevent filter clogging. The maximalcell density was over 10(7) ml(-1), the maximalu-PA activity in supernatant reached 6250IU.ml(-1), and 1604l supernatant containing about51 g u-PA was harvested. Compared to perfusionculture, batch medium-replaced culture could raiseutilizing efficiency of the medium, increase cell specificproductivity and improve the quality of the product which wasnot steady in a 37 degrees C environment. Cells can movefrom seed porous microcarriers occupied by cells tovacant microcarriers spontaneously, withouttrypsinization, and continue to grow until all microcarriers contained cells. It shows that Cytoporeporous microcarriers are very useful and convenient toscale up cultivation step by step.
Hu
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Department of Cell Engineering, Institute of Biotechnology, 20 Dongdajie, Fengtai, Beijing, 100071, P.R. China, hu.xianwen@mailcity.com.
Xiao
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Huang
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Guo
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Zhang
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Li
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