Zhou J, Zhu Y, Mei Q
Department of Physiology and Neuroendocrinology, Fourth Military Medical University, Xian.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 1997 May;13(2):191-2.
A method for primary cell culture of fetal rat gastric fundic epithelial cells was developed. The tissue was incubated with 0.125% trypsin at 4 degrees C for 8-10 hours. The epithelial cells isolated were then cultured in DMEM/F12 medium supplemented with 20% fetal calf serum. Within 24 hours the cells attached to the culture plate and became confluent in 3 days. On phase contrast microscopy, over 90% of cell possessed epithelial characteristics. Immunocytochemical studies showed: (a) 90% of cells were positive in anti-cytokeratin antibody staining; (b) 90% of the epithelial cells contained PAS positive granules; (c) 20% of epithelial cells gave a strong reaction for succinic dehydrogenase activity. Electron microscopy (EM) showed microvilli on the surface of cells, junctional complexes (tight juntion and desmosome), glycogen and mitochondria. Autoradiographic studies showed that these cells possessed the capability to synthesize DNA and this ability was maximum on day 2. This in vitro system may provide a valuable model for studies of cellular functions of gastric mucosa.
建立了一种原代培养胎鼠胃底上皮细胞的方法。将组织在4℃下用0.125%胰蛋白酶孵育8 - 10小时。然后将分离出的上皮细胞培养于补充有20%胎牛血清的DMEM/F12培养基中。细胞在24小时内贴附于培养板，并在3天内汇合。相差显微镜观察显示，超过90%的细胞具有上皮细胞特征。免疫细胞化学研究表明：(a) 90%的细胞抗细胞角蛋白抗体染色呈阳性；(b) 90%的上皮细胞含有PAS阳性颗粒；(c) 20%的上皮细胞琥珀酸脱氢酶活性反应强烈。电子显微镜（EM）显示细胞表面有微绒毛、连接复合体（紧密连接和桥粒）、糖原和线粒体。放射自显影研究表明这些细胞具有合成DNA的能力，且在第2天这种能力最强。该体外系统可为研究胃黏膜细胞功能提供有价值的模型。
