title: Growth, metabolic activity, and productivity of immobilized and freely suspended CHO cells in perfusion culture.
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 × 10(8) 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.
authors.affiliation: School of Chemical & Bioprocess Engineering, University College Dublin, Belfield, Dublin, Ireland; Faculty of Engineering, UAE University, Al Ain, United Arab Emirates.
authors.forename: Ali
authors.initials: A
authors.lastname: Hilal-Alnaqbi
authors.forename: Alan Y C
authors.initials: AY
authors.lastname: Hu
authors.forename: Zhibing
authors.initials: Z
authors.lastname: Zhang
authors.forename: Mohamed
authors.initials: M
authors.lastname: Al-Rubeai
mesh_terms: D000818:Animals; D001692:Biological Transport; D019149:Bioreactors; D016466:CHO Cells; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002453:Cell Cycle; D049109:Cell Proliferation; D018914:Cells, Immobilized / Q000166:cytology / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D007700:Kinetics; D010100:Oxygen / Q000378:metabolism; D010477:Perfusion; D016062:Porosity; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013535:Suspensions; D001616:beta-Galactosidase / Q000096:biosynthesis
chemical_list: D011994:Recombinant Proteins; D013535:Suspensions; D001616:beta-Galactosidase; D010100:Oxygen
keywords: 3D culture; CHO cells; cell cycle; microcarriers; oxygen uptake rate; perfusion culture