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目的培养成人正常食管上皮细胞,建立能够体外长期培养的食管上皮细胞系,为上皮细胞的体外研究提供实验材料。方法取食管癌患者正常食管上皮,用0.25%Dispase酶和0.25%胰蛋白酶/0.02%EDTA消化获取成人食管上皮细胞,使用无血清角化细胞培养液培养,通过细胞形态学观察和角蛋白、上皮膜抗原(EMA)免疫组织化学染色鉴定细胞。结果原代培养8d后,细胞汇合成片呈铺路石样生长,细胞角蛋白、上皮膜抗原表达阳性,可连续传代。结论为体外分离培养成人正常食管上皮细胞建立了方便可行的方法。
Objective To cultivate normal human esophageal epithelial cells and establish long-term culture of esophageal epithelial cell lines in vitro to provide experimental materials for epithelial cells in vitro study. Methods The normal esophageal epithelium of esophageal cancer patients was obtained. The adult esophageal epithelial cells were obtained by digestion with 0.25% Dispase and 0.25% trypsin / 0.02% EDTA, cultured in serum-free keratinocyte culture medium and observed by morphological observation and keratin Cell membrane antigen (EMA) immunohistochemical staining of cells. Results After primary culture for 8 days, the cells were confluent into paving stones, and the positive expression of cytokeratin and epithelial membrane antigen could be continuously passaged. Conclusion The establishment of a convenient and feasible method for the isolation and culture of normal human esophageal epithelial cells in vitro.