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This article is part of the supplement: 22nd European Society for Animal Cell Technology (ESACT) Meeting on Cell Based Technologies

Open Access Meeting abstract

Bag-based rapid and safe seed-train expansion method for Trichoplusia ni suspension cells

Nicole C Bögli1*, Christoph Ries1, Irina Bauer1, Thorsten Adams2, Gerhard Greller2, Regine Eibl1 and Dieter Eibl1

  • * Corresponding author: Nicole C Bögli

Author Affiliations

1 Institute of Biotechnology, Zurich University of Applied Sciences and Facility Management (ZHAW), Wädenswil CH-8820, Switzerland

2 Sartorius Stedim Biotech, Göttingen, D-37075, Germany

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BMC Proceedings 2011, 5(Suppl 8):P124  doi:10.1186/1753-6561-5-S8-P124

The electronic version of this article is the complete one and can be found online at: http://www.biomedcentral.com/1753-6561/5/S8/P124


Published:22 November 2011

© 2011 Bögli et al; licensee BioMed Central Ltd.

This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Meeting abstract

Trichoplusia ni suspension cells (High Five™) used in conjunction with the baculovirus vector expression system (BEVS) are regarded as potential product system of new, recombinant virus-like particle (VLP) vaccines. In order to push vaccine development and production, biomanufacturers use single-use technology when- and wherever possible. This applies to upstream processing and in particular seed-train expansion ranging from cryopreserved vials via t-flasks, spinners (respectively shake flasks) to stirred stainless steel bioreactors. The stainless steel bioreactors deliver inoculum for seed bioreactors and have been increasingly replaced by wave-mixed single-use bag bioreactors during the last 5 years [1].

The approach presented for seed-train cell expansion of High Five suspension cells is based on the Biostat CultiBag RM50 optical (Sartorius Stedim Biotech). It was used for the production of cells for long-term storage and for the expansion of cells for subsequent production experiments. For long-term storage the cells were frozen at high cell concentrations (20 - 40 x 106 cells x mL-1) in 60 mL Cryobags and stored in nitrogen at -196 °C in vapour phase.

Initial experiments were aimed at the growth characterization of High Five suspension cells from a vial working cell bank (WCB). The High Five cells were grown in batch mode and in 250 mL single-use shake flasks (Corning and Sartorius Stedim Biotech) on a Certomat® CT Plus shaker (Sartorius Stedim Biotech) during six days (triplicates, 27 °C, 100 rpm, 25 mm shaking diameter). Afterwards a procedure was developed in which thawed cells from a Cryobag were directly transferred into and expanded in a Biostat CultiBag RM. Under optimal process conditions (500 mL starting volume, a starting cell density of 1 x 106 cells x mL-1, 27 °C, rocking angle of 6 °, 20 - 30 rpm, 0.2 vvm, DO set point 50%) growth rate (0.039 - 0.042 h-1), doubling time (18 - 20 h) and maximal cell density (7.8 - 8.9 x 106 cells x mL-1) showed good correlation with results arising from CultiBags which were inoculated with cells from shake flasks. This bag-based seed-train expansion allows time saving of about one week and reduces cross-contamination, both advantages being due to omitted intermediate cultivation steps in shake flasks.

References

  1. Eibl R, Löffelholz C, Eibl D: Single-Use Bioreactors-An Overview.

    In Single-Use Technology in Biopharmaceutical Manufacture 1st edition. Edited by Eibl R, Eibl D. Wiley. 2010, 1:33-51. OpenURL