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Abstract

ABSTRACT:The aim of this study is to establish the new method of small intestinal epithelial cell culture in order to make intestinal epithelial cell sheets. Cell tissue engineering and cell sheet are very unique technology for cell culture.In the field of regenerative medicine, clinical studies have already attempted to use this technology. In our study, proper cell culture condition for small intestinal cells and the method of making the cell sheet were examined. Conventional intestinal cell culture method was the three-dimensional culture. Cultured cells were unique form and called spheroid.However, this form was not appropriate for making the cell sheet. Therefore, new cell culture condition which was called the two-dimensional culture was necessary to make cell sheets. In our study, cell culture method for using rLaminin-521 coating dish and the mouse embryonic fibroblast which was co-cultured with cultured intestinal cells were the proper way to make small intestinal cell sheets. Indeed, cultured cells under two-dimensional method adhered and stretched on the dish for horizontal direction for two weeks. Gene expressions of culture cells showed that they proved to survive under that condition by confirming the stem cell markers. As to the intestinal cell sheets, the method of co-cultured with MEF enabled the making of small intestinal cell sheet. In conclusion, establishment of small intestinal cell culture in two-dimensional method and making of the intestinal cell sheet. Cell sheets will enable us to provide the efficient cell therapy from its features.