Orbital shaker technology for the cultivation of mammalian cells in suspension.
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 x 10(6) cells/mL for HEK-293 EBNA cells and 5 x 10(6) cells/mL for CHO-DG44 cells in comparison to (2.5-4) x 10(6) 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.
Muller
Natalie
N
Swiss Federal Institute of Technology, Institute of Biological Engineering and Biotechnology, Laboratory of Cellular Biotechnology, Lausanne, Switzerland.
Girard
Philippe
P
Hacker
David L
DL
Jordan
Martin
M
Wurm
Florian M
FM
D000818:Animals; D001709:Biotechnology / Q000191:economics / Q000379:methods*; D018929:Cell Culture Techniques / Q000191:economics / Q000379:methods*; D002478:Cells, Cultured; D015870:Gene Expression; D006801:Humans; D013535:Suspensions; D014162:Transfection
D013535:Suspensions