Perfusion culture of Glycyrrhiza inflata suspension cells in a stir-tank bioreactor
Guirong Wang A and Nian Min Qi B CA School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, P.R. China.
B School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, P.R. China.
C Corresponding author. Email: biotech@sjtu.edu.cn
Australian Journal of Botany 57(3) 240-246 https://doi.org/10.1071/BT08187
Submitted: 15 October 2008 Accepted: 14 April 2009 Published: 29 June 2009
Abstract
Cell and tissue browning is a major impediment of plant culture in vitro. Glycyrrhiza inflata Batalin cells died of lethal browning on Day 9 when cultivated in batch mode. Perfusion culture in bioreactors has a high potential for overcoming plant browning in vitro. In the present study, a continuous perfusion-culture system of G. inflata suspension cells was established in a stir-tank bioreactor. The key component of the bioreactor is an annular gravitational settling zone, through which we achieved continuous separation of cells from the medium. We determined the cell-retention efficiency of the reactor by varying rates of agitation, perfusion and aeration. Complete cell retention was obtained under the following conditions: agitation 0–200 rpm, aeration 0–0.6 volume per volume per minute (vvm) and perfusion rate 0–1.0 day−1. These results were also in good agreement with theoretical estimation. An orthogonal test, including three factors and three levels, showed the optimal culture parameters were as follows: agitation rate 150 rpm, aeration rate 0.3 vvm and perfusion rate 1.0 day−1. Under these conditions, the perfusion-culture system in the bioreactor was operated smoothly, and the maximum cell concentration reached 26 ± 1 g L−1 dry weight.
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