Growth, metabolic activity, and productivity of immobilized and freely suspended CHO cells in perfusion culture

Research output: Contribution to journalArticlepeer-review

Authors

Colleges, School and Institutes

External organisations

  • University College Dublin
  • United Arab Emirates University
  • School of Chemical and Bioprocess Engineering
  • Faculty of Çorlu Engineering
  • University College Dublin
  • UAE University
  • University of Birmingham

Abstract

Chinese hamster ovary (CHO) cells producing β-galactosidase (β-gal) were successfully cultured on silicone-based porous microcarriers (ImmobaSil FS) in a 1 L stirred-tank perfusion bioreactor. We studied the growth, metabolism, and productivity of free and immobilized cells to understand cellular activity in immobilized conditions. CHO cells attached to ImmobaSil FS significantly better than to other microcarriers. Scanning electron microscope images showed that the CHO cells thoroughly colonized the porous surfaces of the ImmobaSil FS, exhibiting a spherical morphology with microvilli that extended to anchorage cells on the silicone surface. In perfusion culture, the concentration of the attached cells reached 8 × 108 cells/mL of carrier, whereas those that remained freely suspended reached 2 × 10 7 cells/mL medium. The β-gal concentration reached more than 5 unit/mL in perfusion culture, more than fivefold that of batch culture. The maximum concentration per microcarrier was proportional to the initial cell density. The specific growth rate, the specific β-gal production rate, the percentage of S phase, and the oxygen uptake rate were all relatively lower for immobilized cells than freely suspended cells in the same bioreactor, indicating that not only do cells survive and grow to a greater extent in a free suspension state, but they are also metabolically more active than viable cells inside the pores of the microcarriers.

Details

Original languageEnglish
Pages (from-to)436-445
Number of pages10
JournalBiotechnology and Applied Biochemistry
Volume60
Issue number4
Publication statusPublished - Jul 2013

Keywords

  • 3D culture, cell cycle, CHO cells, microcarriers, oxygen uptake rate, perfusion culture