Publication Details (including relevant citation information):
Authors: Vishal Gaurav, Susan C. Roberts
Publication date: 2010
Conference name: The American Chemical Society (ACS) Proceedings
Publisher: The ACS National Meeting, San Francisco, CA
Plant cell cultures provide an important route for supply of various natural products including anti-cancer agent paclitaxel. However, cultures are aggregated and highly heterogeneous, with significant cell-to-cell variability in terms of growth and metabolite-production. Flow cytometry is a rapid, powerful and high-throughput tool to investigate and optimize these cultures for enhanced productivity. A novel flow-cytometric method for immunostaining and isolating live high paclitaxel-producing subpopulations from Taxus cell cultures was developed. These cultures were aseptically sorted as high- and low-paclitaxel producing subpopulations using novel florescenceactivated cell-sorting (FACS) technology. Methods to propagate suspension cultures from these isolated subpopulations were developed. Sorted suspension cultures were scaled-up, and their growth, viability, and paclitaxel accumulation analyzed over time. Culture conditions (seeding density, elicitation, media composition - sucrose, growth-hormones, anti-oxidants) were optimized for growth and production. This methodology, through cyto-selection and propagation of high-producers, is an efficient and economical productionmethod for paclitaxel and other plant-derived natural products.