Abstract
AbstractPlant cell cultures of Taxus provide the most reliable production methods for the anti-cancer drug paclitaxel. In order to comprehend the inherent culture heterogeneity and production variability in cell cultures, it is essential that the cellular metabolism is studied at the genomic level. Genomic stability in plant cell cultures is crucial as it affects cell growth and division, metabolite accumulation and protein synthesis. A rapid and efficient method to prepare nuclei suspensions from aggregated cell cultures of Taxus was employed. Methods were subsequently developed to simultaneously stain them for DNA and protein content using Propidium Iodide and Fluorescein Isothiocyanate respectively. Flow cytometry was used to analyze and quantify the DNA content and genome size of Taxus using known reference species as standards. Furthermore, their genomic stability was evaluated by correlating DNA content and genome size with cell size and complexity, protein content, and elicitation effects using multiparameter flow cytometry. These techniques to evaluate and correlate various culture characteristics can be very useful in designing superior bio processes for enhanced production.
Highlights
ObjectivesTo characterize culture heterogeneity through investigation at genomic levelsTo assess the metabolic stability of plant species and understand the progression of cell cycle and DNA content distribution and its control in greater detail through evaluation of DNA content / Genome size and protein contentTo help establish relationships between primary and secondary metabolism to determine the position of the cell within the cell cycle to predict its further development trajectories SampleTo explain how and under what conditions (DNA content, protein content, cell size, metabolic state, etc.) does the mechanisms governing genomic and metabolic stability get activated ApproachRapid and efficient preparation of nuclei suspensions from aggregated plant cell culturesEstablishing simultaneous staining protocols for DNA and protein contentMultiparameter flow cytometric analysis Laser Detectors
In order to comprehend the inherent culture heterogeneity and production variability in cell cultures, it is essential that the cellular metabolism is studied at the genomic level
Genomic stability in plant cell cultures is crucial as it affects cell growth and division, metabolite accumulation and protein synthesis
Summary
ObjectivesTo characterize culture heterogeneity through investigation at genomic levelsTo assess the metabolic stability of plant species and understand the progression of cell cycle and DNA content distribution and its control in greater detail through evaluation of DNA content / Genome size and protein contentTo help establish relationships between primary and secondary metabolism to determine the position of the cell within the cell cycle to predict its further development trajectories SampleTo explain how and under what conditions (DNA content, protein content, cell size, metabolic state, etc.) does the mechanisms governing genomic and metabolic stability get activated ApproachRapid and efficient preparation of nuclei suspensions from aggregated plant cell culturesEstablishing simultaneous staining protocols for DNA and protein contentMultiparameter flow cytometric analysis Laser Detectors. Plant cell cultures of Taxus provide the most reliable production methods for the anti-cancer drug paclitaxel. Genomic stability in plant cell cultures is crucial as it affects cell growth and division, metabolite accumulation and protein synthesis. A rapid and efficient method to prepare nuclei suspensions from aggregated cell cultures of Taxus was employed.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
