An air-liquid interface model of human lung epithelium generated from bronchiolar epithelial cells proliferated using medium containing ROCK kinase inhibitor
10.3969/j.issn.2095-4344.2015.28.028
- VernacularTitle: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
- Author:
Yuanyuan JIA
;
Jinxi HE
;
Yingfei SUN
;
Fei HAN
;
Jiali YANG
;
Yong LI
;
Xiaoming LIU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2015;(28):4582-4587
- CountryChina
- Language:Chinese
-
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.
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.
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.
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.
