Large-Scale Transient Transfection of Chinese Hamster Ovary Cells in Suspension

We describe a one-liter transfection of suspension-adapted Chinese hamster ovary (CHO-DG44) cells using polyethyleneimine (PEI) for DNA delivery. The method involves transfection at a high cell density (5 × 10 cells/mL) by direct addition of plasmid DNA (pDNA) and PEI to the culture and subsequent i...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 1603; p. 45
Main Authors Rajendra, Yashas, Balasubramanian, Sowmya, Hacker, David L
Format Journal Article
LanguageEnglish
Published United States 01.01.2017
Subjects
Animals
Cell Culture Techniques - methods
CHO Cells
Cricetinae
Cricetulus
DNA - administration & dosage
DNA - chemistry
DNA - isolation & purification
Plasmids - administration & dosage
Plasmids - chemistry
Plasmids - isolation & purification
Polyethyleneimine - chemistry
Transfection - methods
Polyethyleneimine
Transfection
Recombinant protein
CHO cells
Orbital shaking
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Summary:We describe a one-liter transfection of suspension-adapted Chinese hamster ovary (CHO-DG44) cells using polyethyleneimine (PEI) for DNA delivery. The method involves transfection at a high cell density (5 × 10 cells/mL) by direct addition of plasmid DNA (pDNA) and PEI to the culture and subsequent incubation at 31 °C with agitation by orbital shaking. We also describe an alternative method in which 90% of the pDNA is replaced by nonspecific (filler) DNA, and the production phase is performed at 31 °C in the presence of 0.25% N, N-dimethylacetamide (DMA).
ISSN:1940-6029
DOI:10.1007/978-1-4939-6972-2_3