Growth kinetics of animal cells immobilized within porous support particles in a perfusion culture

Abstract

The growth kinetics of anchorage-independent animal cells [mouse myeloma MPC-11 (ATCC CCL 167) immobilized within porous polyvinyl formal resin biomass support particles (BSPs; 3 × 3 × 3 mm cubes; mean pore diameter, 60 µm) was analyzed by measuring intracellular and extracellular lactate dehydrogenase (LDH) activities in a perfusion culture. First, the intracellular LDH content of cells immobilized within the BSPs was assayed in a shake-flask culture and found to remain almost comparable to that of non-immobilized cells in the exponential growth phase under static culture. Then, cells inoculated in the BSPs were grown in a continuous stirred-tank bioreactor. Using the LDH content of non-immobilized cells, the density of immobilized cells and the numbers of leaked and dead cells were evaluated, respectively, by the intracellular LDH activity of immobilized and leaked cells and the LDH activity in cell-free culture supernatant. In the initial period, immobilized cells exhibited exponential growth at a constant apparent specific growth rate; however, the acutal specific growth rate, which takes into consideration cell death and cell leakage, decreased significantly. In the stationary phase, the actual specific growth rate maintained a constant but markedly lower value than during exponential growth.