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

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

  • Rajendra Y Laboratory of Cellular Biotechnology (LBTC), École Polytechnique Fédérale de Lausanne (EPFL), CH-1015, Lausanne, Switzerland.
  • Balasubramanian S Laboratory of Cellular Biotechnology (LBTC), École Polytechnique Fédérale de Lausanne (EPFL), CH-1015, Lausanne, Switzerland.
  • Hacker DL Laboratory of Cellular Biotechnology (LBTC), École Polytechnique Fédérale de Lausanne (EPFL), CH-1015, Lausanne, Switzerland. david.hacker@epfl.ch.
  • 2017-05-12
Published in:
  • Methods in molecular biology (Clifton, N.J.). - 2017
CHO cells
Orbital shaking
Polyethyleneimine
Recombinant protein
Transfection
Animals
CHO Cells
Cell Culture Techniques
Cricetinae
Cricetulus
DNA
Plasmids
Polyethyleneimine
Transfection
English 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 × 106 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).
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/177222
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