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Surface immobilization of anchorage‐dependent mammalian cells
Author(s) -
Robert Joëlle,
Côté Johanne,
Archambault Jean
Publication year - 1992
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260390702
Subject(s) - laboratory flask , bioreactor , cell culture , hela , chemistry , cell , tissue culture , matrix (chemical analysis) , biophysics , microbiology and biotechnology , chromatography , materials science , biology , biochemistry , in vitro , genetics , organic chemistry
Anchorage‐dependent HeLa cells were successfully cultured on two fibrous materials (A07 and R100) with porosities of 75–125 and 40 μm, void fractions of 92% and 81%, and fiber diameters of 7.6 and 10.2 μm, respectively, in 100‐mL spinner flasks and 2‐L stirred tank bioreactors. The matrix was formed into a fixed vertical spiral configuration. All cultures displayed rapid (≤2–3 h) attachment of inoculated cells (≥95%) to the matrix, uniform coverage of the immobilizing area with viable cells, and no significant amount of cell debris in the medium. Spinner flask cultures indicated that the denser material R100 showed better results in terms of final cell density. The growth of HeLa cells on material R100 in both culture systems was similar to that observed in tissue culture dishes (specific growth rate ∼0.03–0.04 h −1 , maximum cell density of 8 × 10 6 –9 × 10 6 cells · mL −1 , and yields of 0.4 × 10 8 cells · m M −1 on glucose and 2 × 10 8 −3 × 10 8 cells · m M −1 on glutamine). Scale‐up of this culture technique in a 2‐L bioreactor under perfusion with pH and dissolved oxygen (DO) control yielded cell densities of up to 1.6 × 10 6 cells · mL −1 . Two other anchorage‐dependent mammalian cells (ADC) known to be cultured with difficulty in roller bottles or with micro carriers were easily grown on material R100 in spinner flasks. The performance of this culture technique was compared to other ADC culture systems.