A Method for Culturing Embryonic <em>C. elegans</em> Cells

Abstract

C. elegans is a powerful model system, in which genetic and molecular techniques are easily applicable. Until recently though, techniques that require direct access to cells and isolation of specific cell types, could not be applied in C. elegans. This limitation was due to the fact that tissues are confined within a pressurized cuticle which is not easily digested by treatment with enzymes and/or detergents. Based on early pioneer work by Laird Bloom, Christensen and colleagues 1 developed a robust method for culturing C. elegans embryonic cells in large scale. Eggs are isolated from gravid adults by treatment with bleach/NaOH and subsequently treated with chitinase to remove the eggshells. Embryonic cells are then dissociated by manual pipetting and plated onto substrate-covered glass in serum-enriched media. Within 24 hr of isolation cells begin to differentiate by changing morphology and by expressing cell specific markers. C. elegans cells cultured using this method survive for up 2 weeks in vitro and have been used for electrophysiological, immunochemical, and imaging analyses as well as they have been sorted and used for microarray profiling. Video LinkThe video component of this article can be found at

Mechanosensitive unpaired innexin channels in C. elegans touch neurons

“…Given the challenges of patch-clamp recordings in vivo in C. elegans, we continued our characterization of the mechanosensitive channel in cultured touch neurons (56). Embryonically derived ALM and PLM touch neurons are present in culture, represent ϳ0.5% of the population of cells, and differentiate recapitulating the properties they have in vivo (7,56).…”

“…Embryonically derived ALM and PLM touch neurons are present in culture, represent ϳ0.5% of the population of cells, and differentiate recapitulating the properties they have in vivo (7,56). Postembryonic neurons AVM and PVM are not present in culture.…”

“…elegans cell culture. C. elegans embryonic cells were cultured following a published procedure (15,56). Briefly, large quantities of gravid adults were grown on enriched peptone plates (8P plates) seeded with Escherichia coli strain Na22.…”

Knock-out of a mitochondrial sirtuin protects neurons from degeneration in Caenorhabditis elegans

“…Caenorhabditis elegans embryonic cells, including touch neurons, can be dissociated and cultured in in vitro where they differentiate and express cell specific markers [ 29 , 61 , 62 ]. We cultured Caenorhabditis elegans embryonic cells from wild type, mec-4(d) , and mec-4(d);sir-2 .…”

“…Embryonic cell culture was performed as described previously [ 62 ]. Briefly, a large number of gravid adult worms were grown on enriched peptone agar plates (8P) with NA22 Escherichia Coli , collected in 50 ml tubes, washed 3 times with sterile H 2 O and centrifuged at 1200 rpm for 10 minutes.…”