Yamada Kiyoshi, Sato Kanako, Morishita Satoru, Kaminogawa Shuichi, Totsuka Mamoru
Department of Applied Biological Chemistry, The University of Tokyo, Japan.
Biosci Biotechnol Biochem. 2009 Aug;73(8):1849-55. doi: 10.1271/bbb.90246. Epub 2009 Aug 7.
Studies of the physiological functions of intestinal epithelial cells (IECs) have been limited by the difficulty of primary culture of IEC. We established a method for primary culture of mouse IEC by culturing fragments of fetal small intestines pretreated with EDTA. This method reproducibly resulted in the expansion of cytokeratin-positive epithelial cells, and vigorous expansion of the epithelial cells was observed only from intestinal fragments of embryonic days 15-16. These cells expressed alkaline phosphatase activity and major histocompatibility complex (MHC) class II molecules, indicating the mature phenotype of IEC in a small intestine. The cells also presented antigens to CD4(+) T cells. Furthermore, the cells expressed various cytokines and chemokines, and the expression was enhanced by bacterial stimulation. These results indicate that the primary-cultured mouse IEC prepared by the method established here can be a beneficial tool in study of the functions of IECs, especially in mucosal immunity.
肠道上皮细胞（IECs）生理功能的研究一直受到IEC原代培养困难的限制。我们通过培养经乙二胺四乙酸（EDTA）预处理的胎儿小肠片段，建立了一种小鼠IEC原代培养方法。该方法可重复性地导致细胞角蛋白阳性上皮细胞的扩增，并且仅在胚胎第15 - 16天的肠道片段中观察到上皮细胞的旺盛扩增。这些细胞表达碱性磷酸酶活性和主要组织相容性复合体（MHC）II类分子，表明小肠中IEC的成熟表型。这些细胞还向CD4(+) T细胞呈递抗原。此外，这些细胞表达多种细胞因子和趋化因子，并且细菌刺激可增强其表达。这些结果表明，通过此处建立的方法制备的原代培养小鼠IEC可成为研究IEC功能，特别是黏膜免疫功能的有益工具。
