中国组织工程研究Issue(28)：4582-4587,6.DOI:10.3969/j.issn.2095-4344.2015.28.028
人细支气管上皮细胞的ROCK激酶抑制剂培养体系扩增及气液相模型的建立
An air-liquid interface model of human lung epithelium generated from bronchiolar epithelial cells proliferated using medium containing ROCK kinase inhibitor
摘要
Abstract
BACKGROUND:Primary human lung epithelial cel s are difficult to be isolated and cultured in vitro, which is characterized as limited sources, low cel viability, slow proliferation capacity, and lacking of differentiation capability. <br> OBJECTIVE:To establish an air-liquid interface model of lung epithelium by in vitro proliferation of human bronchiolar epithelial cel s, which is used for research on function of lung epithelial cel s. <br> METHODS:Primary human bronchiolar epithelial cel s were isolated using Pronase and DNase I combined digestive methods, and then proliferated using medium containing ROCK kinase inhibitor. The proliferated cel s were used for establishment of the air-liquid interface epithelium model. Cel differentiation was identified using scanning electron microscope, phase contrast microscope and immunofluorescent staining. <br> RESULTS AND CONCLUSION:The primary human bronchiolar epithelial cel s could be expanded successful y using medium containing ROCK kinase inhibitor, and the basal cel marker Cytokeratin14 was preferential y expressed in the proliferated cel population, indicating that these basal cel s might be the main subpopulation of human lung epithelial stem cel s. Subsequently, the proliferated cel s under the air-liquid interface could differentiate into ciliated cel s and non-ciliated column cel s. The results suggest that the proliferation and differentiation of human bronchiolar epithelial cel s were maintained in the presence of ROCK kinase inhibitor, and the air-liquid interface could promote the differentiation of human bronchiolar epithelial cel s.关键词
干细胞/培养/人细支气管上皮细胞/肺脏/ROCK激酶抑制剂/气液相培养/国家自然科学基金分类
医药卫生引用本文复制引用
贾元元,何进喜,孙颖飞,韩飞,杨佳丽,李勇,刘晓明..人细支气管上皮细胞的ROCK激酶抑制剂培养体系扩增及气液相模型的建立[J].中国组织工程研究,2015,(28):4582-4587,6.基金项目
国家自然科学基金(31172278)@@@@Funding:the National Natural Science Foundation of China, No.31172278 （31172278）
