Vishal Gaurav - Plant cell culture process development: Use of Flow Cytometry for creation of specialized cell lines

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  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.

  Address (URL): Use_of_Flow_Cytometry_for_creation_of_specialized_cell_lines.pdf