Liquid-culture protocols for synchronous starvation, growth, dauer formation, and dietary restriction of Caenorhabditis elegans

Abstract

Standard laboratory culture of Caenorhabditis elegans utilizes solid growth media with a bacterial food source. However, this culture method limits control of food availability and worm population density, factors that impact many life-history traits. Here, we describe liquid-culture protocols for precisely modulating bacterial food availability and population density, facilitating reliable production of arrested L1 larvae, dauer larvae, dietarily restricted worms, or well-fed worms. Worms can be grown in small quantities for standard assays or in the millions for other applications. For complete details on the use and execution of these protocols, please refer to Hibshman et al. (2016), Webster et al. (2018), and Jordan et al. (2019).

Keywords: Cell biology; Genetics; Model organisms.

Graphical abstract

Figure 1

Representative images of C. elegans arrested as L1 larvae and dauer larvae resulting from liquid-culture growth in Applications I and III (A) Starved L1 larvae. (B) Dauer larvae. Scale bars, 50 μm.

Figure 2

Representative images of C. elegans fed ad libitum or restricted diets in Application IV (A) Adult worm after 96 h in ad libitum culture (25 mg/mL HB101). (B) Adult worm after 96 h in dietary restriction (3.1 mg/mL HB101). Scale bars, 200 μm.