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

• Published in: Plant cell, tissue and organ culture Vol. 117; no. 3; pp. 393 - 399
• Main Authors: Havenith, Heide, Raven, Nicole, Di Fiore, Stefano, Fischer, Rainer, Schillberg, Stefan
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer-Verlag 01.06.2014; Springer Netherlands; Springer Nature B.V
• Subjects: Antibodies; Biomedical and Life Sciences; cell suspension culture; cells; Cultivation; Culture media; Cytology; Fluorescence; Fluorescence microscopy; humans; Image processing; Life Sciences; Mammalian cells; Mammals; Microorganisms; Monoclonal antibodies; Morphology; Nicotiana tabacum; Original Paper; Plant cells; Plant Genetics and Genomics; plant morphology; Plant Pathology; Plant Physiology; Plant Sciences; Productivity; recombinant proteins; Tobacco; Transgenic plants; Zebrafish; Automated cell counting; Growth rate determination; Molecular farming; Transgenic BY-2 cells; OPERA; Human monoclonal antibody
• ISSN: 0167-6857; 1573-5044
• DOI: 10.1007/s11240-014-0448-x

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.