An air-liquid interface model of human lung epithelium generated from bronchiolar epithelial cells proliferated using medium containing ROCK kinase inhibitor

doi:10.3969/j.issn.2095-4344.2015.28.028

Abstract

Abstract:

BACKGROUND: Primary human lung epithelial cells are difficult to be isolated and cultured in vitro, which is characterized as limited sources, low cell 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 cells, which is used for research on function of lung epithelial cells.
METHODS: Primary human bronchiolar epithelial cells were isolated using Pronase and DNase I combined digestive methods, and then proliferated using medium containing ROCK kinase inhibitor. The proliferated cells were used for establishment of the air-liquid interface epithelium model. Cell differentiation was identified using scanning electron microscope, phase contrast microscope and immunofluorescent staining.
RESULTS AND CONCLUSION: The primary human bronchiolar epithelial cells could be expanded successfully using medium containing ROCK kinase inhibitor, and the basal cell marker Cytokeratin14 was preferentially expressed in the proliferated cell population, indicating that these basal cells might be the main subpopulation of human lung epithelial stem cells. Subsequently, the proliferated cells under the air-liquid interface could differentiate into ciliated cells and non-ciliated column cells. The results suggest that the proliferation and differentiation of human bronchiolar epithelial cells were maintained in the presence of ROCK kinase inhibitor,
and the air-liquid interface could promote the differentiation of human bronchiolar epithelial cells.
BACKGROUND: Primary human lung epithelial cells are difficult to be isolated and cultured in vitro, which is characterized as limited sources, low cell 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 cells, which is used for research on function of lung epithelial cells.
METHODS: Primary human bronchiolar epithelial cells were isolated using Pronase and DNase I combined digestive methods, and then proliferated using medium containing ROCK kinase inhibitor. The proliferated cells were used for establishment of the air-liquid interface epithelium model. Cell differentiation was identified using scanning electron microscope, phase contrast microscope and immunofluorescent staining.
RESULTS AND CONCLUSION: The primary human bronchiolar epithelial cells could be expanded successfully using medium containing ROCK kinase inhibitor, and the basal cell marker Cytokeratin14 was preferentially expressed in the proliferated cell population, indicating that these basal cells might be the main subpopulation of human lung epithelial stem cells. Subsequently, the proliferated cells under the air-liquid interface could differentiate into ciliated cells and non-ciliated column cells. The results suggest that the proliferation and differentiation of human bronchiolar epithelial cells were maintained in the presence of ROCK kinase inhibitor,
and the air-liquid interface could promote the differentiation of human bronchiolar epithelial cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Lung, Bronchioles, Epithelial Cells

CLC Number:

R394.2

Jia Yuan-yuan, He Jin-xi, Sun Ying-fei, Han Fei, Yang Jia-li, Li Yong, Liu Xiao-ming. An air-liquid interface model of human lung epithelium generated from bronchiolar epithelial cells proliferated using medium containing ROCK kinase inhibitor [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(28): 4582-4587.

Figures/Tables 4

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