Back
Low-temperature pausing of cultivated mammalian cells.
Journal article

Low-temperature pausing of cultivated mammalian cells.

Show more…
  • 2004-12-08
Published in:
  • Biotechnology and bioengineering. - 2005
Animals
Bioreactors
CHO Cells
Cell Culture Techniques
Cell Line
Cell Proliferation
Cell Survival
Cricetinae
Cricetulus
Cryopreservation
Humans
Kidney
Temperature
Time Factors
English There are currently two methods for maintaining cultured mammalian cells, continuous passage at 37 degrees C and freezing in small batches. We investigated a third approach, the "pausing" of cells for days or weeks at temperatures below 37 degrees C in a variety of cultivation vessels. High cell viability and exponential growth were observed after pausing a recombinant Chinese hamster ovary cell line (CHO-Clone 161) in a temperature range of 6-24 degrees C in microcentrifuge tubes for up to 3 weeks. After pausing in T-flasks at 4 degrees C for 9 days, adherent cultures of CHO-DG44 and human embryonic kidney (HEK293 EBNA) cells resumed exponential growth when incubated at 37 degrees C. Adherent cultures of CHO-DG44 cells paused for 2 days at 4 degrees C in T-flasks and suspension cultures of HEK293 EBNA cells paused for 3 days at either 4 degrees C or 24 degrees C in spinner flasks were efficiently transfected by the calcium phosphate-DNA coprecipitation method, yielding reporter protein levels comparable to those from nonpaused cultures. Finally, cultures of a recombinant CHO cell line (CHO-YIgG3) paused for 3 days at 4 degrees C, 12 degrees C, or 24 degrees C in bioreactors achieved the same cell mass and recombinant protein productivity levels as nonpaused cultures. The success of this approach to cell storage with rodent and human cell lines points to a general biological phenomenon which may have a wide range of applications for cultivated mammalian cells.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/78003
Statistics
Document views: 63 File downloads: