关键词: 胃壁细胞;细胞分离;细胞培养

    摘 要:目的 完成胃壁细胞分离及纯化，建立原代培养胃壁细胞的方法. 方法 制备大鼠翻转的胃囊用含链霉蛋白酶E的消化液注入胃囊内孵育，并用磁力搅拌器轻轻搅拌而制备单个的胃底腺细胞，Percoll梯度离心富集胃壁细胞，用含100mL·L-1 血清的PBS或培养液，时差贴壁去除成纤维细胞，然后，将壁细胞接种于培养板中，培养液用无血清的1∶1Harm's F-12/DMEM培养，内含胰岛素5mg·L-1 ，氢化可地松4μg·L-1 ，转铁蛋白5mg·L-1 ，硒酸钠5μg·L-1 ，牛血清白蛋白2g·L-1 ，表皮生长因子25μg·L-1 ，葡萄糖1.98g·L-1 持续培养可达1wk以上. 结果 获得的壁细胞经吖啶橙(acridine orange，AO)鉴定纯度达80%以上，原代培养经HE染色可见壁细胞呈分散小组式生长，且生长状态良好. 结论 此方法适宜于大鼠胃壁细胞的分离及体外培养，为体外进一步研究壁细胞的功能打下基础.

    Isolation and primary culture of gastric parietal cells of rats

    CHEN Lei HUANG Wei-Quan SUN Xu-De LüBao-Zhen PU Ruo-Lei

    1 Deparment of Histology and Embryology，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China，

    2 Department of Anesthesiology，Tangdu Hospital，Fourth Military Medical University，Xi'an710038，China

    Keywords:gastric parietal cell;cell isolation;cell culture

    Abstract:AIM To establish a new model of isolation and primary culture in vitro for further study.METHODS The stomachs of rats were reverted to form sacs with the mucosa facing outward and serosa facing inward.Pronase E solution was injected into the sac lumina.The stomachs were incubat-ed in950mL·L-1 O2 and50mL·L-1 CO2 at37℃for60min ......