Differentiated cultures of primary hamster tracheal airway epithelial cells |
| |
Authors: | Regina?K?Rowe Steven?L?Brody Email author" target="_blank">Andrew?PekoszEmail author |
| |
Institution: | (1) Molecular Microbiology and Microbial Pathogenesis Graduate Program, Washington University School of Medicine, 660 South Euclid Avenue, Campus Box 8230, 63110 St. Louis, Missouri;(2) Department of Molecular Microbiology, Washington University School of Medicine, 660 South Euclid Avenue, Campus Box 8230, 63110 St. Louis, Missouri;(3) Department of Medicine, Washington University School of Medicine, 660 South Euclid Avenue, Campus Box 8230, 63110 St. Louis, Missouri;(4) Department of Pathology and Immunology, Washington Univeristy School of Medicine, 660 South Euclid Avenue, Campus Box 8230, 63110 St. Louis, Missouri |
| |
Abstract: | Summary Primary airway epithelial cell cultures can provide a faithful representation of the in vivo airway while allowing for a controlled
nutrient source and isolation from other tissues or immune cells. The methods used have significant differences based on tissue
source, cell isolation, culture conditions, and assessment of culture purity. We modified and optimized a method for generating
tracheal epithelial cultures from Syrian golden hamsters and characterized the cultures for cell composition and function.
Soon after initial plating, the epithelial cells reached a high transepithelial resistance and formed tight junctions. The
cells differentiated into a heterogeneous, multicellular culture containing ciliated, secretory, and basal cells after culture
at an air-liquid interface (ALI). The, secretory cell populations initially consisted of MUC5AC-positive goblet cells and
MUC5AC/CCSP double-positive cells, but the makeup changed to predominantly Clara cell secretory protein (CCSP)-positive Clara
cells after 14 d. The ciliated cell populations differentiated rapidly after ALI as judged by the appearance of β tubulin
IV-positive cells. The cultures produced mucus, CCSP, and trypsin-like proteases and were capable of wound repair as judged
by increased expression of matrilysin. Our method provides an efficient, high-yield protocol for producing differentiated
hamster tracheal epithelial cells that can be used for a variety of in vitro studies including tracheal cell differentiation,
airway disease mechanisms, and pathogen-host interactions. |
| |
Keywords: | goblet Clara respiratory virus trachea influenza wound repair ribonucleic acid virus |
本文献已被 PubMed SpringerLink 等数据库收录! |
|