Controlling Heterogeneity in Plant Cell Culture and Secondary Product Accumulation through Metabolic Analysis and Modeling of Population Behavior

通过代谢分析和群体行为建模控制植物细胞培养和次级产物积累中的异质性。具体目标是：（1）使用多参数流式细胞术获得基线红豆杉种群信息以表征代谢异质性，并开发种群平衡方程模型以将代谢数据与工艺规模信息相关联，（2）使用新的荧光激活细胞分选（FACS）技术基于紫杉醇积累分离亚群，并表征亚群中的代谢异质性，以及（3）基于紫杉醇积累对亚群进行分类和恢复，分析生长和紫杉醇积累方面的长期稳定性，并开发具有预测群体行为能力的群体平衡方程模型。智力优势：从拟议的研究结果将提供一个详细的表征红豆杉异质性，然后可以用来设计有效的策略，以稳定和优化紫杉醇积累。这是非常有价值的，因为未来紫杉醇的供应很可能在很大程度上取决于植物细胞培养技术的使用。更广泛的影响：这项研究的结果有可能对科学界创造新方法产生重大影响（例如，单细胞分析和单植物细胞的分选）和研究红豆杉和其他植物系统的研究人员的方法。所有参与该项目的人员都将接受实验和建模方法的培训，并将积极参与一个新的指导计划。 代表性不足的学生将通过NSF赞助的教育计划招募。

Controlling Heterogeneity in Plant Cell Culture and Secondary Product Accumulation through Metabolic Analysis and Modeling of Population BehaviorSusan RobertsUniversity of MassachusettsCBET-0730779The 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.