Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Food & Nutrition Science Regular Papers
Establishment of a Primary Culture Method for Mouse Intestinal Epithelial Cells by Organ Culture of Fetal Small Intestine
Kiyoshi YAMADAKanako SATOSatoru MORISHITAShuichi KAMINOGAWAMamoru TOTSUKA
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2009 Volume 73 Issue 8 Pages 1849-1855

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Abstract

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.

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© 2009 by Japan Society for Bioscience, Biotechnology, and Agrochemistry
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