Optimisation of growth conditions for ovine airway epithelial cell differentiation at an air-liquid interface

Autor: Erin Sutherland, Robert L. Davies, Catherine C. Berry, Nicky O’Boyle
Jazyk: angličtina
Rok vydání: 2018
Předmět:
0301 basic medicine
Cellular differentiation
Cell
Retinoic acid
lcsh:Medicine
Epithelium
chemistry.chemical_compound
Epidermal growth factor
Animal Cells
Immunofluorescence Staining
Medicine and Health Sciences
lcsh:Science
Cells
Cultured
Epithelial cell differentiation
Staining
Multidisciplinary
Chemistry
Air
Cell Staining
Cell Differentiation
Cell biology
Trachea
medicine.anatomical_structure
Physical Sciences
Models
Animal
Cellular Types
Anatomy
Porosity
Cell Division
Research Article
Chemical Elements
Histology
030106 microbiology
Primary Cell Culture
Stratified squamous epithelium
Tretinoin
Research and Analysis Methods
03 medical and health sciences
Species Specificity
medicine
Animals
Cilia
Sheep
Dose-Response Relationship
Drug
Epidermal Growth Factor
lcsh:R
DAPI staining
Biology and Life Sciences
Epithelial Cells
Cell Biology
Culture Media
Oxygen
030104 developmental biology
Biological Tissue
Microscopy
Fluorescence
Cell culture
Specimen Preparation and Treatment
Nuclear staining
Microscopy
Electron
Scanning
lcsh:Q
Developmental Biology
Zdroj: PLoS ONE
PLoS ONE, Vol 13, Iss 3, p e0193998 (2018)
ISSN: 1932-6203
Popis: Respiratory tract infections are of significant concern in the agriculture industry. There is a requirement for the development of well-characterised in vitro epithelial cell culture models in order to dissect the diverse molecular interactions occurring at the host-pathogen interface in airway epithelia. We have analysed key factors that influence growth and differentiation of ovine tracheal epithelial cells in an air-liquid interface (ALI) culture system. Cellular differentiation was assessed at 21 days post-ALI, a time-point which we have previously shown to be sufficient for differentiation in standard growth conditions. We identified a dose-dependent response to epidermal growth factor (EGF) in terms of both epithelial thickening and ciliation levels. Maximal ciliation levels were observed with 25 ng ml-1 EGF. We identified a strict requirement for retinoic acid (RA) in epithelial differentiation as RA exclusion resulted in the formation of a stratified squamous epithelium, devoid of cilia. The pore-density of the growth substrate also had an influence on differentiation as high pore-density inserts yielded higher levels of ciliation and more uniform cell layers than low pore-density inserts. Differentiation was also improved by culturing the cells in an atmosphere of sub-ambient oxygen concentration. We compared two submerged growth media and observed differences in the rate of proliferation/expansion, barrier formation and also in terminal differentiation. Taken together, these results indicate important differences between the response of ovine tracheal epithelial cells and other previously described airway epithelial models, to a variety of environmental conditions. These data also indicate that the phenotype of ovine tracheal epithelial cells can be tailored in vitro by precise modulation of growth conditions, thereby yielding a customisable, potential infection model.
Databáze: OpenAIRE
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