Cultivation of mammalian cells as aggregates in bioreactors: Effect of calcium concentration of spatial distribution of viability

Madhusudan V. Peshwa, Yun‐Seung ‐S Kyung, Don B. McClure, Wei‐Shou ‐S Hu

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61 Scopus citations

Abstract

Recombinant human kidney epithelial 293 cells were cultivated as aggregates in suspension. The concentration calcium ion, in the range of 100 μM to 1mM, 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 lower viability in the interior. With its ease of retention in a perfusion bioreactor, aggregate cultures offer an alternative choice for large‐scale operation. © 1993 John Wiley & Sons, Inc.

Original languageEnglish (US)
Pages (from-to)179-187
Number of pages9
JournalBiotechnology and bioengineering
Volume41
Issue number2
DOIs
StatePublished - Jan 20 1993

Keywords

  • aggregates
  • animal cell bioreactor
  • confocal microscopy
  • mammalian cells

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