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大鼠前脑分化原代神经元的高通量转染

High-throughput transfection of differentiated primary neurons from rat forebrain.

作者信息

Marine Shane, Freeman Jamie, Riccio Antonella, Axenborg Marie-Louise, Pihl Johan, Ketteler Robin, Aspengren Sara

机构信息

Department of Automated Biotechnology, Merck & Co., Inc., North Wales, PA, USA.

出版信息

J Biomol Screen. 2012 Jun;17(5):692-6. doi: 10.1177/1087057112439233. Epub 2012 Mar 8.

Abstract

Primary neurons in culture are considered to be a highly relevant model in the study of neuronal development and activity. They can be cultivated and differentiated in vitro but are difficult to transfect using conventional methods. To address this problem, a capillary electroporation system called Cellaxess Elektra was developed for efficient and reproducible transfection of primary cortical and hippocampal neurons without significant impact on cell morphology and viability. The cells are transfected in any stage of differentiation and development, directly in cell culture plates. Genetic material is delivered in situ to as many as 384 samples at a time, which enables both high-throughput and high-quality screening for hard-to-transfect primary cells, meaning that gene function can be studied on a genome-wide scale in cells previously inaccessible to genetic manipulation.

摘要

培养的原代神经元被认为是神经元发育和活动研究中高度相关的模型。它们可以在体外培养和分化,但使用传统方法难以转染。为了解决这个问题,开发了一种名为Cellaxess Elektra的毛细管电穿孔系统,用于高效且可重复地转染原代皮质和海马神经元,而对细胞形态和活力没有显著影响。细胞在分化和发育的任何阶段,直接在细胞培养板中进行转染。遗传物质可一次原位递送至多达384个样本,这使得对难以转染的原代细胞进行高通量和高质量筛选成为可能,也就是说,可以在以前无法进行基因操作的细胞中进行全基因组规模的基因功能研究。

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