Choosing the Right Differentiation Medium to Develop Mucociliary Phenotype of Primary Nasal Epithelial Cells In Vitro.
Cell Differentiation
/ drug effects
Cells, Cultured
Culture Media
/ pharmacology
Enzyme-Linked Immunosorbent Assay
Epidermal Growth Factor
/ metabolism
Epithelial Cells
/ cytology
Humans
Immunohistochemistry
Microscopy, Electron, Scanning
Nasal Mucosa
/ cytology
Tretinoin
/ pharmacology
Vascular Endothelial Growth Factor A
/ metabolism
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
24 04 2020
24 04 2020
Historique:
received:
21
01
2020
accepted:
06
04
2020
entrez:
26
4
2020
pubmed:
26
4
2020
medline:
15
12
2020
Statut:
epublish
Résumé
In vitro differentiation of airway epithelium is of interest for respiratory tissue engineering and studying airway diseases. Both applications benefit from the use of primary cells to maintain a mucociliated phenotype and thus physiological functionality. Complex differentiation procedures often lack standardization and reproducibility. To alleviate these shortfalls, we compared differentiation behavior of human nasal epithelial cells in four differentiation media. Cells were differentiated at the air-liquid interface (ALI) on collagen-coated inserts. Mucociliary differentiation status after five weeks was analyzed by electron microscopy, histology and immunohistochemistry. The amount of ciliation was estimated and growth factor concentrations were evaluated using ELISA. We found that retinoic-acid-supplemented mixture of DMEM and Airway Epithelial Cell Growth Medium gave most promising results to obtain ciliated and mucus producing nasal epithelium in vitro. We discovered the balance between retinoic acid (RA), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and fibroblast growth factor β (FGF-β) to be relevant for differentiation. We could show that low VEGF, EGF and FGF-β concentrations in medium correspond to absent ciliation in specific donors. Therefore, our results may in future facilitate donor selection and non-invasive monitoring of ALI cultures and by this contribute to improved standardization of epithelial in vitro culture.
Identifiants
pubmed: 32332878
doi: 10.1038/s41598-020-63922-8
pii: 10.1038/s41598-020-63922-8
pmc: PMC7181704
doi:
Substances chimiques
Culture Media
0
Vascular Endothelial Growth Factor A
0
Tretinoin
5688UTC01R
Epidermal Growth Factor
62229-50-9
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
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