幽门螺杆菌感染新型原代人胃上皮细胞模型的建立

目的 培养三维(3D)人胃黏膜上皮类器官,转为二维(2D)原代胃黏膜细胞培养,并建成人2D原代胃上皮细胞的幽门螺杆菌感染模型。 方法 (1)从正常人胃上皮组织中分离胃腺,在含有多种生长调节和凋亡抑制等混合因子的培养基中,依附于基质胶而培养成3D类器官；(2)利用免疫荧光技术鉴定胃上皮类器官的相关分子标记；(3)研究正常原代胃上皮细胞被幽门螺杆菌感染后的形态学变化,利用免疫印迹技术鉴定幽门螺杆菌感染相关蛋白的表达水平。 结果 成功培养出可长期传代的人胃上皮3D类器官,具有典型的人胃黏膜上皮分子标记。而且3D类器官转为2D平面培养的原代胃上皮细胞,可作为幽门螺杆菌的体外原代细胞感染模型。 结论 3D胃上皮类器官,可作为2D原代胃上皮细胞的持久来源,为研究幽门螺杆菌感染人体胃上皮的分子机制带来个体化的新模型。

Establishment of human gastric primary cell culture system for Helicobacter pylori infection in vitro

Objective To cultivate 3D human gastric epithelial organoids, and transfer into 2D primary gastric epithelial cells, which could be used as infection model for Helicobacter pylori (H.pylori). Methods (1) Gastric glands were isolated from normal gastric epithelial tissues, and were further grown into gastric organoid on the matrigel with the medium containing various growth factors and apoptosis inhibitors. (2) Immunofluorescence was used to identify relevant molecular markers of gastric epithelial organoids. (3) The morphological changes of normal primary gastric epithelial cells infected with H.pylori were observed, and the expression of H.pylori infection related proteins were identified using western blot. Results The 3D human gastric organoids were obtained successfully, which could be passaged for a long time and had typical molecular markers of human gastric epithelium. In addition, 2D cultured primary gastric epithelial cells from 3D organoids could be used as H.pylori infection model in vitro. Conclusion 3D gastric organoids can be used as a constant source of 2D primary gastric epithelial cells, which provide an individualized new primary cell model for research on the molecular mechanism of H.pylori infection in vitro.