Bangladesh Vet J (2010) 44(1-4):12-20 *Corresponding author: Telephone: +880 91 67401~6/ Ext. 2330; Fax: +880 91 61510; Cellphone: +88-01713815104; e-mail: shafiqpharma@yahoo.co.uk Copyright © Bangladesh Veterinary Association All right reserved 0030-9915/10 Isolation and Characterization of Intestinal Sub-epithelial Myofibroblast in Rat: An in vitro Tool for Bioresearch System M. S. Islam 1,2* , H. Ozaki 2 1 Department of Pharmacology, Faculty of Veterinary Science, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh. 2 Department of Veterinary Pharmacology, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Tokyo, Japan. ABSTRACT Primary cell culture is a valuable tool that extensively used in the basic research of organ specific functions, physiology, pathology, pharmacology and so on. The aim of this study was to derive primary mesenchyme cell line representative of colon tissues. First, we identified the location of sub-epithelial myofibroblast in the colon tissues in rat. -smooth muscle-actin (- SMA) positive myofibroblast found just beneath the epithelia of colon tissues. Accordingly, we isolated intestinal sub-epithelial myofibroblasts (ISEMFs) from colonic lamina propria of rat. The isolated cells were cultured and characterized by phase contrast, immunofluorescent and RT-PCR analysis. The harvested cells expressed the specific markers such as -SMA, vimentin and were negative for desmin expression. Finally, we did trans-well migration assay to know the difficulties or convenience of this isolated cell culture experiment. The isolated rat ISEMFs grow well in culture media and easy to culture for the research purposes. Therefore, the primary ISEMF might be a useful alternative in vitro tool for bioresearch study. Key words: Intestinal sub-epithelial myofibroblast, -SMA, Colon, Rat INTRODUCTION Sub-epithelial myofibroblasts are located at the interface between the epithelium and lamina propria in most mucosal tissues (Valentich et al., 1997). Ultrastructural features of ISEMFs demonstrate both smooth muscle cell and fibroblasts and are thought to be intermediate cell of smooth muscle cell and fibroblast cells (Andoh et al., 2005). Although the cells have been characterized in many ways but still the physiological roles of ISMFs