Establishment of a Primary Culture Method for Mouse Intestinal Epithelial Cells by Organ Culture of Fetal Small Intestine
Establishment of a Primary Culture Method for Mouse Intestinal Epithelial Cells by Organ Culture of Fetal Small Intestine
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DOI：
10.1271/bbb.90246
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发表时间：
2009-08-01
影响因子：
1.6
通讯作者：
Totsuka, Mamoru
中科院分区：
文献类型：
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作者：
Yamada, Kiyoshi;Sato, Kanako;Totsuka, Mamoru
肠上皮细胞原代培养的困难限制了对肠上皮细胞生理功能的研究。采用EDTA预处理的胎鼠小肠组织块，建立了原代培养小鼠IEC的方法。该方法可重复地导致细胞角蛋白阳性上皮细胞的扩增，并且仅从胚胎第15-16天的肠碎片中观察到上皮细胞的剧烈扩增。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IEC的成熟表型。这些细胞还将抗原呈递给CD 4（+）T细胞。此外，细胞表达多种细胞因子和趋化因子，并且通过细菌刺激增强表达。结果表明，本方法制备的原代培养小鼠IEC可作为研究IEC功能，特别是粘膜免疫功能的有效工具。
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.