A primary culture system for functional analysis of C-elegans neurons and muscle cells
A primary culture system for functional analysis of C-elegans neurons and muscle cells
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DOI：
10.1016/s0896-6273(02)00591-3
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发表时间：
2002-02-14
期刊：
影响因子：
16.2
通讯作者：
Strange, K
中科院分区：
文献类型：
--
作者：
Christensen, M;Estevez, A;Strange, K
线虫为研究神经肌肉系统的功能和发育提供了重要的见解。然而，动物的体积很小，限制了进入单个神经元和肌肉细胞进行生理、生化和分子研究。我们在这里描述了使线虫胚胎细胞在体外分化为神经元和肌肉细胞的原代培养方法。形态、电生理和GFP报告研究表明，培养细胞的分化和功能特性与体内观察到的相似。通过荧光激活的细胞分选可以产生表达特定GFP报告的丰富的细胞群体。在培养基中加入双链RNA可以显著抑制靶基因的表达。线虫细胞原代培养为该领域迄今没有的各种实验机会提供了新的基础。
C. elegans has provided important insights into neuromuscular system function and development. However, the animal's small size limits access to individual neurons and muscle cells for physiological, biochemical, and molecular study. We describe here primary culture methods that allow C. elegans embryonic cells to differentiate into neurons and muscle cells in vitro. Morphological, electrophysiological, and GFP reporter studies demonstrate that the differentiation and functional properties of cultured cells are similar to those observed in vivo. Enriched populations of cells expressing specific GFP reporters can be generated by fluorescence-activated cell sorting. Addition of double-stranded RNA to the culture medium induces dramatic knockdown of targeted gene expression. Primary nematode cell culture provides a new foundation for a wide variety of experimental opportunities heretofore unavailable in the field.