Culture of embryonic C. elegans cells for electrophysiological and pharmacological analyses.

Abstract

Despite the considerable advantages that C. elegans offers for studying gene function in vivo, this system is quite challenging for in vivo electrophysiological analysis of channel function, particularly in neurons. A major problem is that C. elegans neurons are confined in a pressurized and hard-to-penetrate cuticle. Recently, a method for culturing C. elegans embryonic cells has been developed and numerous researchers have already applied this option to study a variety of native ion channels and transporters using various configurations of the patch-clamp technique. C. elegans embryonic cells are obtained from eggs harvested from synchronized gravid adults and then are dissociated using a combination of enzymatic treatment and manual pipetting. Once plated on a surface covered with peanut lectin, cells adhere and differentiate into neurons, muscle and epithelial cells. Cultured embryonic cells recapitulate the expression of differentiation markers and are found in the culture in proportion to their cell type in the mature embryo. Differentiated cells survive well for at least 2 weeks. It should be noted that postembryonic cells do not appear to be generated in these cultures. Cultures can be used for electrophysiological study, testing of pharmacological sensitivities, and for RNAi. C. elegans cell culture thus constitutes the basis for the application of experimental procedures that are not easily applicable to the intact nematode.