Fox C C, Dvorak A M, Peters S P, Kagey-Sobotka A, Lichtenstein L M
J Immunol. 1985 Jul;135(1):483-91.
Study of the role of mast cells in the human gastrointestinal tract has suffered from the inability to examine these cells in vitro. In addition, work in rodent systems suggests that there are substantial differences between intestinal mast cells and those from other tissues, making extrapolation of data from other sources difficult. We report a method for producing mast cell-containing single cell suspensions from human intestinal tissue by mechanical and enzymatic dispersion. This method yields 4.5 X 10(5) mast cells per gram of tissue in purity of 3.1 +/- 2.1%. These mast cells were functionally intact as assessed by survival in short-term culture, low spontaneous release, and appropriate IgE-mediated histamine release. They were morphologically intact on electron microscopy and conformed to published descriptions of human lung mast cells. The intestinal mucosal mast cells were also indistinguishable from human lung mast cells in histamine content, goat anti-human IgE dose-response curves, kinetics of histamine release, unresponsiveness to f-met peptide, and production of arachidonic acid metabolites, prostaglandin D2, and leukotriene C4. This procedure produces human intestinal mast cell suspensions in sufficient numbers to make pharmacologic characterization and further purification of this cell feasible.
对人类胃肠道中肥大细胞作用的研究一直受到无法在体外检测这些细胞的困扰。此外，啮齿动物系统的研究表明，肠道肥大细胞与其他组织的肥大细胞存在显著差异，这使得从其他来源推断数据变得困难。我们报告了一种通过机械和酶分散从人肠道组织中产生含肥大细胞的单细胞悬液的方法。该方法每克组织可产生4.5×10⁵个肥大细胞，纯度为3.1±2.1%。通过短期培养中的存活、低自发释放以及适当的IgE介导的组胺释放评估，这些肥大细胞功能完整。电子显微镜下它们形态完整，符合已发表的人肺肥大细胞描述。肠道黏膜肥大细胞在组胺含量、山羊抗人IgE剂量反应曲线、组胺释放动力学、对f-甲硫氨酸肽无反应以及花生四烯酸代谢产物、前列腺素D2和白三烯C4的产生方面也与人肺肥大细胞无区别。该程序可产生足够数量的人肠道肥大细胞悬液，使对该细胞进行药理学表征和进一步纯化成为可能。
