Primary culture of Caenorhabditis elegans developing embryo cells for electrophysiological, cell biological and molecular studies.

Primary culture of Caenorhabditis elegans developing embryo cells for electrophysiological, cell biological and molecular studies. Research Abstract Details 

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Primary culture of Caenorhabditis elegans developing embryo cells for electrophysiological, cell biological and molecular studies. Abstract Text:

Kevin Strange,Michael Christensen,Rebecca Morrison,Kevin Strange,Michael Christensen,Rebecca Morrison,

Cell culture is an invaluable tool for investigation of basic biological processes. However, technical hurdles including low cell yield, poor cell differentiation and poor attachment to the growth substrate have limited the use of this tool for studies of the genetic model organism Caenorhabditis elegans. This protocol describes a method for the large-scale culture of C. elegans embryo cells. We also describe methods for in vitro RNA interference, fluorescence-activated cell sorting of embryo cells and imaging of cultured cells for patch-clamp electrophysiology studies. Developing embryos are isolated from gravid adult worms. After eggshell removal by enzymatic digestion, embryo cells are dissociated and plated onto glass substrates. Isolated cells terminally differentiate within 24 h. Analysis of gene expression patterns and cell-type frequency suggests that in vitro embryo cell cultures recapitulate the developmental characteristics of L1 larvae. Cultured embryo cells are well suited for physiological analysis as well as molecular and cell biological studies. The embryo cell isolation protocol can be completed in 5-6 h.

Primary culture of Caenorhabditis elegans developing embryo cells for electrophysiological, cell biological and molecular studies. Publishing Authors By Initials

K Strange,M Christensen,R Morrison,K Strange,M Christensen,R Morrison,

For similar genetic processes: gene expression regulation: epigenesis, genetic: gene silencing: rna interference research abstracts see: genetic processes: gene expression regulation: epigenesis, genetic: gene silencing: rna interference research

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Primary culture of Caenorhabditis elegans developing embryo cells for electrophysiological, cell biological and molecular studies. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Nature protocols

VOLUME: 2

Page Numbers: 1003-12

Journal Abbreviation:

ISSN: 1750-2799

DAY: 3

MONTH: 12

YEAR: 2007

Primary culture of Caenorhabditis elegans developing embryo cells for electrophysiological, cell biological and molecular studies. Information

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LANGUAGE: eng

NlmUniqueID: 101284307

Primary culture of Caenorhabditis elegans developing embryo cells for electrophysiological, cell biological and molecular studies. Keywords Mesh Terms:

KEYWORDS: RNA Interference

MESH TERMS: physiology

Chemical & Substance for Abstract: Primary culture of Caenorhabditis elegans developing embryo cells for electrophysiological, cell biological and molecular studies. Information

Substance Name: Green Fluorescent Proteins

Registry Number: 147336-22-9

Grant and Affiliation Information for Primary culture of Caenorhabditis elegans developing embryo cells for electrophysiological, cell biological and molecular studies.

AFFILIATION: Department of Anesthesiology, Vanderbilt University Medical Center, Nashville, Tennessee 37232, USA. kevin.strange@vanderbilt.edu

Country: England

AGENCY: United States NIDDK

GRANT: R01 DK51610

ACRONYM: DK

MEDLINETA: Nat Protoc

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