CULTIVATION OF MAMMALIAN-CELLS AS AGGREGATES IN BIOREACTORS - EFFECT OF CALCIUM-CONCENTRATION ON SPATIAL-DISTRIBUTION OF VIABILITY

Recombinant human kidney epithelial 293 cells were cultivated as aggregates in suspension. The concentration of calcium ion, in the range of 100 muM to 1 mM, affected the rate of aggregate formation. During the course of cultivation the size distribution of aggregates shifted and the fraction of larger aggregates increased. This effect was more profound in cultures with a high calcium concentration. Scanning and transmission microscopic examination of the aggregates revealed that cell packing was greater in the high calcium cultures and that ultrastructural integrity was retained in aggregates from both low and high calcium cultures. Confocal microscopy was applied to examine the viability of cells in the interior of the aggregates. High viability was observed in the aggregates obtained from exponentially growing cultures. Aggregates from the high calcium culture in the stationary phase exhibited a lower viability in the interior. With its ease of retention in a perfusion bioreactor, aggregate cultures offer an alternative choice for large-scale operations.