A new culture system for the cultivation of mammalian cells for the production of several biologically active substances.

We recently developed a new dialysis culture system (termed LIFROC-device) for the cultivation of lymphokine-activated killer cells (Murata et al., 1990, 1991). In the present study, we applied the LIFROC-device (400 ml culture vessel) to the cultivation of mammalian cells for the production of biologically active substances. We cultured mouse-mouse hybridoma TP-709, secreting anti-tisse plasminogen activator (tPA) monoclonal antibody (mAb), recombinant CHO GT19, secreting hGH, and human melanoma Bowes cells, secreting tPA. With the LIFROC-device, TP-709 grew to a maximal cell density of 3.8 x 10(6) cells/ml and and produced 480 micrograms/ml (192 mg in total) of mAb. GT19 reached a cell density of 2.2 x 10(6) cells/ml and produced 302 micrograms/ml (120 mg in total) of hGH. Bowes cells expanded to 4.4 x 10(6) cells/ml and secreted 8.5 micrograms/ml (3.3 mg in total) of tPA. The protein concentration in the culture broths of the LIFROC-device became 7-200 times higher than that of batch culture. Thus, the LIFROC-device can be applied to protein production as well as cell growth with high efficiency.