Abstract
A cell-detaching reactor was developed to collect cells growing on microcarriers for inoculation between stepwise-expanded bioreactors. It consisted of a trypsinization zone and a separation zone, which were separated by a 200-mesh stainless steel screen. The screen allowed the cells only to pass through to the next bioreactor, after the cells have been trypsinized and detached from microcarriers. The operating feasibility of the cell-detaching reactor was tested with anchorage-dependent recombinant Chinese hamster ovary (rCHO) and African green monkey kidney (Vero) cells. rCHO and Vero cells were first cultured in a small microcarrier bioreactor, and then inoculated via the cell-detaching reactor into either a packed-bed bioreactor (for rCHO cells) or a larger microcarrier bioreactor (for Vero cells). For rCHO cells, the cell density reached 1.3 × 107 cells/ml in the perfusion culture, and Vero cells reached 1.3 × 106 cells/ml in the batch culture.
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Acknowledgements
We thank Mr. Weimin Fan and Mr. Jian Lu for their assistance in the setup of the cell-detaching reactor, and Dr Yanjun Liu for his kind providing rCHO cells. We also thank Dr Huiching Hia and Dr Sijing Wang for critical reading of this manuscript. This work was partially funded by the Project of Chinese National Programs for High Technology Research and Development (2002AA217011).
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Sun, X., Zhang, Y. A cell-detaching reactor for inoculation of anchorage-dependent CHD and Vero cells between stepwise-expanded bioreactors. Biotechnol Lett 29, 697–701 (2007). https://doi.org/10.1007/s10529-006-9294-1
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DOI: https://doi.org/10.1007/s10529-006-9294-1