Nutrient regulation by continuous feeding for large-scale expansion of mammalian cells in spheroids

Bradley P. Weegman, Ahmad Essawy, Peter Nash, Alexandra L. Carlson, Kristin J. Voltzke, Zhaohui Geng, Marjan Jahani, Benjamin B. Becker, Klearchos K. Papas, Meri T. Firpo

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

In this demonstration, spheroids formed from the β-TC6 insulinoma cell line were cultured as a model of manufacturing a mammalian islet cell product to demonstrate how regulating nutrient levels can improve cell yields. In previous studies, bioreactors facilitated increased culture volumes over static cultures, but no increase in cell yields were observed. Limitations in key nutrients such as glucose, which were consumed between batch feedings, can lead to limitations in cell expansion. Large fluctuations in glucose levels were observed, despite the increase in glucose concentrations in the media. The use of continuous feeding systems eliminated fluctuations in glucose levels, and improved cell growth rates when compared with batch fed static and SSB culture methods. Additional increases in growth rates were observed by adjusting the feed rate based on calculated nutrient consumption, which allowed the maintenance of physiological glucose over three weeks in culture. This method can also be adapted for other cell types.

Original languageEnglish (US)
Article numbere52224
JournalJournal of Visualized Experiments
Volume2016
Issue number115
DOIs
StatePublished - Sep 25 2016

Keywords

  • Bioengineering
  • Bioreactor
  • Continuous feeding
  • Culture
  • Expansion
  • Glucose
  • Issue 115
  • Nutrient regulation
  • Spheroid
  • Stirred suspension bioreactor

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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