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《Chinese Journal of Coll Biology》 1997-01
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STUDIES ON ISOLATION AND PRIMARY CULTURE OF NEONATE RAT INTESTINAL EPITHELIAL CELL

DAI Ding Wei, WU Sheng Mei, LI Min, LIAO Xian Ping, LIU Zai Yong(Shanghai Institute for Pediatric Research, Xinhua Hospital, Shanghai Second MedicalUniversity. Shanghai 200092)  
Trypsin, collagenase Ⅰ and combined dispase (grade Ⅰ) and collagenase Ⅰ or collagenase Ⅺ were used to isolate neonate rat intestinal epithelial cell (IEC) and the results were compared and the optimal culture conditions were determined. We developed the technique of isolation and primary culture of neonate rat IEC. The results showed that satisfactory isolation of the epithelia and good attachment were achieved by using the collagenase Ⅺ dispase (grade Ⅰ) di-gestion. Coating the glass culture flaskes with bovine dermal collagens or rat tail tendon col-lagens I was beneficial to the attachment of IEC in culture. IEC proliferation in vitro is ini-tially dependent upon the factors produced by heterogeneous mesenchymal celis, also critically dependent upon the quality of the culture medium and constituents used. IEC attached within 1-2 days, proliferated obviously at 7-8 days. Cultures reached conflunce within 10-14 days. Immunocytochemical characterization showed that more than 90% attached cells were positive for cytokeratin and epithelial cell membrane antigen. Histochemical staining for alkaline pho-sphatase (AKP) revealed that AKP activity was expressed in more than 90% attached cells. Electronmicroscopy confirmed apical microvilli in IEC. The results provide a very useful expe-rimental model to study physiology and pathology of IEC.
【Fund】: 上海市高等学校科学技术发展基金
【CateGory Index】: R329
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