title: Culturing Caenorhabditis elegans in axenic liquid media and creation of transgenic worms by microparticle bombardment.
abstract: In this protocol, we present the required materials, and the procedure for making modified C. elegans Habituation and Reproduction media (mCeHR). Additionally, the steps for exposing and acclimatizing C. elegans grown on E. coli to axenic liquid media are described. Finally, downstream experiments that utilize axenic C. elegans illustrate the benefits of this procedure. The ability to analyze and determine C. elegans nutrient requirement was illustrated by growing N2 wild type worms in axenic liquid media with varying heme concentrations. This procedure can be replicated with other nutrients to determine the optimal concentration for worm growth and development or, to determine the toxicological effects of drug treatments. The effects of varied heme concentrations on the growth of wild type worms were determined through qualitative microscopic observation and by quantitating the number of worms that grew in each heme concentration. In addition, the effect of varied nutrient concentrations can be assayed by utilizing worms that express fluorescent sensors that respond to changes in the nutrient of interest. Furthermore, a large number of worms were easily produced for the generation of transgenic C. elegans using microparticle bombardment.
authors.affiliation: Department of Animal and Avian Sciences, University of Maryland.
authors.forename: Tamika K
authors.initials: TK
authors.lastname: Samuel
authors.affiliation: Department of Animal and Avian Sciences, University of Maryland.
authors.forename: Jason W
authors.initials: JW
authors.lastname: Sinclair
authors.affiliation: Department of Animal and Avian Sciences, University of Maryland.
authors.forename: Katherine L
authors.initials: KL
authors.lastname: Pinter
authors.affiliation: Department of Animal and Avian Sciences, University of Maryland; Department of Cell Biology and Molecular Genetics, University of Maryland; hamza@umd.edu.
authors.forename: Iqbal
authors.initials: I
authors.lastname: Hamza
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D017173:Caenorhabditis elegans / Q000235:genetics* / Q000254:growth & development*; D003470:Culture Media; D006418:Heme; D010316:Particle Size
chemical_list: D003470:Culture Media; D006418:Heme