By Jason D. Brown, Mark W. Signs, Jamie Q. White, PhD, Sridhar Viamajala, PhD, and Kamal A. Rashid, PhD
Volume 9, Issue 2 (Winter 2010/2011)
Nematode worms release pheromones to communicate information about food, stress, and sex to other individuals. The nematode species C. elegans is a genetic system for studying the biology underlying these pheromone-mediated behaviors. Identifying the chemical structure of C. elegans pheromones requires purifying them directly from large quantities of conditioned growth media. Release of pheromones may depend on growth conditions, population density, and developmental stage. However, published studies only report nematode pheromones from developmentally asynchronous cultures at variable population densities. Here, we have developed reliable methods for liquid culture of developmentally-synchronous populations of C. elegans at controlled population densities from 5,000 to 10,000 worms/mL at the scale of 10 to 100 L. Generating synchronous populations requires harvesting eggs from adults using alkaline bleach. In this study, we developed modified tangential flow filtration (TFF) systems to rapidly bleach adults and recover large quantities of healthy, viable eggs. This method achieved synchronous cultures at scales 2.5 x greater than cultures based on agar plates, and 5 x greater than previously reported for synchronous liquid cultures. We routinely harvested 2.5 x 10E7 animals (total) from a single 5 L large-scale culture. Furthermore, our TFF system effectively recovered pheromone-containing media at all scales tested. This work makes it feasible to isolate other biologically useful secreted molecules from transgenic C. elegans…
Citation: Brown JD, Signs MW, White JQ, Viamajala S, Rashid KA. Large-Scale, Developmentally-Synchronous Caenorhabditis elegans Liquid Cultures Using a New Tangential-Flow Filtration Method. BioProcess J, 2011; 9(2): 34-43. https://dx.doi.org/10.12665/J92.Brown
Posted online February 9, 2011