University of Massachusetts CBET-0730779
The proposed research project centers on the use of novel approaches to study and characterize plant cell culture heterogeneity. The specific aims are to: (1) obtain baseline Taxus population information using multiparameter flow cytometry to characterize metabolic heterogeneity and develop population balance equation models to correlate metabolic data with process scale information, (2) isolate subpopulations based on paclitaxel accumulation using novel Fluorescence Activated Cell Sorting (FACS) techniques, and characterize metabolic heterogeneity and in the subpopulations, and (3) sort and recover subpopulations based on paclitaxel accumulation, analyze long-term stability in terms of growth and paclitaxel accumulation, and develop population balance equation models with the ability to predict population behavior.
Intellectual Merit: The results from the proposed research will provide a detailed characterization of Taxus heterogeneity, which can then be used to design effective strategies to stabilize and optimize paclitaxel accumulation. This is of great value because future paclitaxel supply is likely to depend significantly on the use of plant cell culture technology.
Broader Impacts: The results of this research have the potential to significantly impact the scientific community in the creation of new methodologies (e.g., single cell analysis and sorting of single plant cells) and approaches for investigators studying Taxus and other plant systems. All personnel participating in this project will be trained in both experimental and modeling approaches and will actively participate in a novel mentoring program. Underrepresented students will be recruited through the NSF-sponsored educational program.
