Viability characterization of Taxus chinensis plant cell suspension cultures by rapid colorimetric- and image analysis-based techniques

被引:4
|
作者
Wucherpfennig, Thomas [1 ]
Schulz, Annika [1 ]
Pimentel, Jaime Arturo [2 ]
Corkidi, Gabriel [2 ]
Sieblitz, Dominik [3 ]
Pump, Matthias [3 ]
Gorr, Gilbert [3 ]
Schuette, Kai [3 ]
Wittmann, Christoph [1 ]
Krull, Rainer [1 ]
机构
[1] Tech Univ Carolo Wilhelmina Braunschweig, Inst Biochem Engn, D-38106 Braunschweig, Germany
[2] UNAM, Image Anal Lab, Inst Biotechnol, Cuernavaca, Morelos, Mexico
[3] Phyton Biotech GmbH, Ahrensburg, Germany
关键词
Taxus chinensis plant cell culture; Plant cell aggregates; Viability; Aggregate coloration; Alamar Blue assay; Image analysis; TAXOL PRODUCTION; AGGREGATE SIZE; ALAMAR BLUE; 2,3,5-TRIPHENYLTETRAZOLIUM CHLORIDE; MECHANICAL-STRESS; CALLUS-CULTURES; ASSAY; GROWTH; SURVIVAL; BACCATA;
D O I
10.1007/s00449-014-1153-1
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
For the commercially established process of paclitaxel production with Taxus chinensis plant cell culture, the size of plant cell aggregates and phenotypic changes in coloration during cultivation have long been acknowledged as intangible parameters. So far, the variability of aggregates and coloration of cells are challenging parameters for any viability assay. The aim of this study was to investigate simple and non-toxic methods for viability determination of Taxus cultures in order to provide a practicable, rapid, robust and reproducible way to sample large amounts of material. A further goal was to examine whether Taxus aggregate cell coloration is related to general cell viability and might be exploited by microscopy and image analysis to gain easy access to general cell viability. The Alamar Blue assay was found to be exceptionally eligible for viability estimation. Moreover, aggregate coloration, as a morphologic attribute, was quantified by image analysis and found to be a good and traceable indicator of T. chinensis viability.
引用
收藏
页码:1799 / 1808
页数:10
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