|
Author:
Date:
2012-11-20
[Abstract] The gastrointestinal (GI) tract is lined by a single layer of epithelial cells which function in secretion, absorption, and digestion. In addition, most GI tract tumors develop from epithelial cells (carcinomas). This protocol describes isolation of the surface epithelium from the underlying stroma, muscular layer and submucosa in the GI tract. In this protocol, epithelial cell adhesions are weekend by chelating Ca +2 ions followed by mechanical separation of the cells by vortexing. Analysis of protein levels and gene expression patterns in isolated epithelial cells versus whole GI tissue minimizes the potential for confounding contributions from contaminating stromal cells.
[摘要] 胃肠（GI）道由在分泌，吸收和消化中起作用的单层上皮细胞排列。 此外，大多数胃肠道肿瘤从上皮细胞（癌）发展。 该协议描述了表面上皮从下面的基质，肌肉层和胃肠道中的粘膜下层的分离。 在该协议中，上皮细胞粘附是周末通过螯合Ca +2离子，然后通过涡旋机械分离的细胞。 分离的上皮细胞与整个GI组织中蛋白质水平和基因表达模式的分析使来自污染性基质细胞的混杂贡献的潜力最小化。
|