胚胎鼠胃粘膜上皮细胞的原代培养

PRIMARY CULTURE OF FETAL RAT GASTRIC MUCOSAL EPITHELIUM

采用胰酶冷消化法对胚胎鼠胃粘膜上皮细胞进行了培养，细胞培养于含２０％的胎牛血清的ＤＭＥＭ／Ｆ１２的培养液中，种植２４ｈ后开始生长，３ｄ长成片状，相差显微镜下观察９０％的细胞具有上皮细胞特征。免疫细胞化学显示：（ａ）９０％的细胞上皮角蛋白染色阳性；（ｂ）９０％的上皮细胞ＰＡＳ染色阳性；（ｃ）２０％的上皮细胞琥珀酸脱氢酶（ＳＤＨ）染色阳性。透射电镜可见微绒毛、连接复合物（紧密连接，桥粒）、丰富的糖原、线粒体。放射自显影结果表明：上皮细胞具有合成ＤＮＡ的能力，而且在培养２ｄ时增殖能力最强。本实验为研究胃粘膜的功能提供了较好的方法。

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℃ for 8-10 hours. The epithelial cells isolated were then cultured in DMEM/F 12 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 cells 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.