Orbital shaker technology for the cultivation of mammalian cells in suspension
- 23 December 2004
- journal article
- research article
- Published by Wiley in Biotechnology & Bioengineering
- Vol. 89 (4), 400-406
- https://doi.org/10.1002/bit.20358
Abstract
For large-scale applications in biotechnology, cultivation of mammalian cells in suspension is an essential prerequisite. Typically, suspension cultures are grown in glass spinner flasks filled to less than 50% of the nominal volume. We propose a superior system for suspension cultures of mammalian cells based on orbital shaker technology. We found that “square-shaped” bottles (square bottles) provide an inexpensive but efficient means to grow HEK-293 EBNA and CHO-DG44 cells to high density. Cultures in agitated 1-L square bottles exceeded the performance of cultures in spinner flasks, reaching densities up to 7 × 106 cells/mL for HEK-293 EBNA cells and 5 × 106 cells/mL for CHO-DG44 cells in comparison to (2.5–4) × 106 cells/mL for cultures of the same cells grown in spinner flasks. For 1-L square bottles, optimal cell growth and viability were observed with a filling volume of 30–40% of the nominal volume and an agitation speed of 130 rpm at a rotational diameter of 2.5 cm. Transient reporter gene expression following gene delivery by calcium phosphate–DNA co-precipitation was the same or slightly better for HEK-293 EBNA cells grown in square bottles as compared to spinner flasks. Reductions in cost, simplified handling, and better performance in cell growth and viability make the agitated square bottle a new and very promising tool for the cultivation of mammalian cells in suspension.Keywords
This publication has 18 references indexed in Scilit:
- Oxygen transfer by orbital shaking of square vessels and deepwell microtiter plates of various dimensionsBiochemical Engineering Journal, 2004
- A study of oxygen transfer in shake flasks using a non‐invasive oxygen sensorBiotechnology & Bioengineering, 2003
- Improvements in spinner-flask designs for insect-cell suspension cultureBiotechnology and Applied Biochemistry, 2003
- Characterization of gas–liquid mass transfer phenomena in microtiter platesBiotechnology & Bioengineering, 2002
- Impact of cell culture process changes on endogenous retrovirus expressionBiotechnology & Bioengineering, 2002
- Balancing GFP reporter plasmid quantity in large‐scale transient transfections for recombinant anti‐human Rhesus‐D IgG1 synthesisBiotechnology & Bioengineering, 2002
- 100-liter transient transfectionCytotechnology, 2002
- Transient gene expression: Recombinant protein production with suspension‐adapted HEK293‐EBNA cellsBiotechnology & Bioengineering, 2001
- Characterisation of the gas–liquid mass transfer in shaking bioreactorsBiochemical Engineering Journal, 2001
- High Throughput Cultivation of Animal Cells Using Shaken Microplate TechniquesPublished by Springer Science and Business Media LLC ,2001