改良成人胃粘膜上皮细胞原代培养  

Modified Primary Culture of Adult Gastric Mucosal Epithelial Cells

作　　者：李志芳[1] 林庚金[1] 蔡定芳[1] 

机构地区：[1]上海医科大学附属华山医院消化内科,200040

出　　处：《胃肠病学》2000年第1期34-35,共2页Chinese Journal of Gastroenterology

摘　　要：目的：建立成人周粘膜上皮细胞原代培养方法。方法：采用胰酶和胶原酶冷消化法消化分离手术切除的成人胃粘膜，细胞培养于含20％小牛血清的DMEM／F12培养液中。结果：细胞于种植24h后开始生长，3天长成片状，相差显微镜下可见90％以上的细胞具有上皮细胞特征。免疫组化显示90％以上的细胞上皮角蛋白染色、上皮膜抗原阳性。MTT比色法显示活细胞数于第4天达到高峰。3H-胸腺嘧啶核苷（TdR）掺入法显示细胞具有合成DNA的能力，并于第3天达到高峰。结论：本方法培养的细胞特异性高，存活时间长，为体外研究提供了较理想的模型。Background/Aims: To establish a method for primary culture of adult gastric mucosal epithelialcells. Methods: The resected tissue was incubated with trypsin and collagenase Ⅱ at 4℃ for 8~10hours. The isolated epithelial cells were cultured in DMEM/F12 medium supplemented with 20% fetalcalf serum. Results: Within first 24 hours of culture, the cells attached to the plate and became con-fIuent in 3 days. On phase contrast microscopy, over 90% of the cells possessed epithelial characte-ristics. Immunohistochemical studies showed that over 90% of the cells were positive for anti-cyto-keratin and anti-epithelial mucosal antigen-antibody staining. The active cells could be detected pre-dominantly 4 days after MTT staining. The synthesis of DNA was detected by 3H-TdR involvement andreached the peak on the third day. Conclusions: This method system provides a valuable model forstudies of gastric mucosal epithelial cells in vitro.

关 键 词：细胞培养 胃粘膜 上皮细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]