Image-based analysis of cell-specific productivity for plant cell suspension cultures

Plant Cell, Tissue and Organ Culture (PCTOC)(2014)

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摘要
More and more plant cell suspension cultures are regarded as an attractive alternative to mammalian cells as host organism for production of complex recombinant proteins. The most important advantages of the production platform are low costs, easy scalability and enhanced safety by complete lack of animal components in the cultivation media. In order to characterize, understand and control such systems accurately, it is important to determine the cell-specific productivity (Qp) of plant cell-based production platforms. Compared to many microbial and mammalian cells the morphology of plant cells is nonhomogeneous and the cells tend to form aggregates, therefore commercial cell counting systems are too unreliable to determine cell numbers in plant suspension cultures. We addressed this limitation by developing a novel cell counting method based on a combination of cell-staining and automated confocal fluorescence microscopy. This method allowed us, for the first time, to determine the cell-specific productivity of transgenic tobacco ( Nicotiana tabacum cv. Bright Yellow-2) cell suspension cultures producing the human antibody M12. In the future this method will be a useful tool in the development of optimized plant cell-based production processes.
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关键词
Automated cell counting,Growth rate determination,Human monoclonal antibody,Molecular farming,OPERA,Transgenic BY-2 cells
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