人支气管上皮细胞的体外培养

Culture of human bronchial epithelial cells in vitro

背景:人支气管上皮细胞培养在呼吸系统疾病研究中应用越来越广泛。目的:探索美国典型培养物保藏中心人支气管上皮细胞体外培养方法的可行性。方法:对人支气管上皮细胞培养条件进行反复摸索,最终确定应用含体积分数20%胎牛血清的DMEM/F12培养基进行培养。结果与结论:实验培养的人支气管上皮细胞扁平,呈多边形,长满后呈"铺路石"样分布,经免疫组化检测发现培养的细胞表达上皮细胞标志物细胞角蛋白。在支气管扩张症患者痰液上清刺激下,细胞表达的核因子κB、肿瘤坏死因子α和白细胞介素8明显增强,证实培养的人支气管上皮细胞获得成功。提示不必拘泥于美国典型培养物保藏中心推荐的培养条件,改进的培养方法简便易行,有应用价值。

BACKGROUND： Culture of human bronchial epithelial cells has been increasingly applied in the research of respiratory diseases. OBJECTIVE： To explore a feasible in vitro culture method for human bronchial epithelial cell line supplied by American Type Culture Collection. METHODS： Different culture conditions of human bronchial epithelial cells were tested with trials and errors. A feasible method with DMEM/F12 containing 20% fetal bovine serum was finalized. RESULTS AND CONCLUSION： The cultured human bronchial epithelial cells were flat, polygonal, and displayed a cobblestone-like distribution when overgrown. Immunohistochemical staining results showed an expression of epithelial marker keratin in cultured cells. The expression of nuclear factor KB, tumor necrosis factor a and interleukin 8 was increased significantly in cultured cells exposed to sputum supernatant from patients with bronchiectasis. These findings demonstrated that the cultured human bronchial epithelial cells were obtained successfully. Therefore, there is no need to stick to the culture condition recommended by American Type Culture Collection. The modified culture method is simple, feasible and valuable.