A new seed-train expansion method for recombinant mammalian cell lines
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A new approach has been developed and used to minimize the timeand more carefully monitor and control the seed-train expansionprocess of recombinant mammalian cell lines. The process uses 50or 100 ml cryo-bags that contain frozen cells at high cell densities of 20 × 106 ml-1 (100 ml bags) or 40 × 106 cells ml-1 (50 ml bags). The frozen bag cell suspension is thawed and transferred directly into a bioreactorthat has been modified such that pH, DO and temperature can becontrolled at the initial volume of two liters (the working volume eventually increases to 12 l). The successful use of thesecryo-bags and the modified `inoculation' bioreactor to initiate anew seed train expansion of rBHK or rCHO cells is described herein. The interval between cell thawing and the accumulation ofsufficient cell mass to inoculate a production reactor is reducedby at least 25 to 30 days compared to the conventional method that begins with the thaw of 1–2 ml cryo-vials. This `one-step'technology leads to a much more consistent scale-up by reducingmanual operations and avoiding subjective decisions during the scale-up phase. The cell metabolic rates and product integritywere similar to the control experiments. Furthermore, it was found that it is not necessary to include a wash step to removeDMSO prior to the inoculation.
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- A new seed-train expansion method for recombinant mammalian cell lines
Volume 38, Issue 1-3 , pp 99-108
- Cover Date
- Print ISSN
- Online ISSN
- Kluwer Academic Publishers
- Additional Links
- cell banking
- cryo bags
- inoculation bioreactor
- one step inoculation
- seed-train expansion
- Industry Sectors
- Author Affiliations
- 1. Biotechnology Unit, Bayer Corporation, Berkeley, CA, USA
- 2. Bayer AG, D-42117, Wuppertal, Germany