Tompers Dennie M, Labosky Patricia A
University of Pennsylvania, Department of Cell and Developmental Biology, Philadelphia, Pennsylvania 19104-6058, USA.
Stem Cells. 2004;22(3):243-9. doi: 10.1634/stemcells.22-3-243.
The manipulation of embryonic stem (ES) cells to generate targeted mutations via homologous recombination has proved an invaluable resource for researchers from fields as diverse as embryology, immunology, physiology, and biophysics. The derivation and culture of these cells has been described in many places, but there is no substitute for hands-on experience. In lieu of that, here we provide a step-by-step guide for the technique of electroporation, complete with micrographs of the cells at each step of the process. This is intended to allow the first time ES cell worker, as well as the more experienced researcher, to view the morphology of ES cells under ideal conditions as well as when the cells are suboptimal and should be discarded.
通过同源重组操纵胚胎干细胞(ES细胞)以产生靶向突变,已被证明是胚胎学、免疫学、生理学和生物物理学等不同领域研究人员的宝贵资源。这些细胞的衍生和培养在许多地方都有描述,但实践经验无可替代。在此,我们提供电穿孔技术的分步指南,并配有该过程每个步骤的细胞显微照片。这旨在让首次操作ES细胞的人员以及经验更丰富的研究人员,能够在理想条件下以及细胞状态欠佳应丢弃时,观察ES细胞的形态。