Perfusion culture of hybridoma cells using a centrifuge to separate cells from culture mixture

Abstract

The influence of centrifugal force on the growth of cells was examined by exposing the cells of the mouse-human hybridoma H1 line to centrifugal forces (100 g and 500 g) for 30 min twice a day and comparing the static culture with that of unexposed cells. The centrifugal force of 100 g permitted the line to proliferate as in unexposed cases, while 500 g evidently inhibited their proliferation. High density cultivation was done by a perfusion system with the mouse-mouse hybridoma JTC-3 line and the mouse-human hybridoma X87 line where the cells were separated from the culture medium by centrifugation. In the cultivation of the JTC-3 line in a serum-free culture, the viable cell density reached 10 7 cells/ml. This result almost corresponded to that in the cultivation where the cells were separated from the culture medium by gravitation. JTC-3 line was cultured in a 10% FCS-added medium and a serum-free medium, and the maximum viable cell density exceeded 10 7 cells/ml in both of them. In the serum-free culture medium, the cell density and the antibody productivity were found to be almost equal to the cultivation with cells separation from the culture mixture by gravitation.