Choosing the Right Differentiation Medium to Develop Mucociliary Phenotype of Primary Nasal Epithelial Cells In Vitro

Scientific Reports, vol. 10, pp. 6963

Abstract

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.

Authors 7

1. Anja E. Luengen corresponding Aachen Department of Biohybrid and Medical Textiles (BioTex) AME-Institute of Applied Medical Engineering

   Maastricht University · RWTH Aachen University

   Affiliation as printed

   Aachen-Maastricht Institute for Biobased Materials, Faculty of Science and Engineering, Maastricht University, Brightlands Chemelot Campus, 6167 RD, Geleen, The Netherlands. luengen@ame.rwth-aachen.de

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Forckenbeckstraße 55, 52074, Aachen, Germany. luengen@ame.rwth-aachen.de

   Aachen-Maastricht Institute for Biobased Materials, Faculty of Science and Engineering, Maastricht University, Brightlands Chemelot Campus, 6167 RD, Geleen, The Netherlands

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Aachen, Germany

2. Caroline Kniebs Aachen Department of Biohybrid and Medical Textiles (BioTex) AME-Institute of Applied Medical Engineering

   Maastricht University · RWTH Aachen University

   Affiliation as printed

   Aachen-Maastricht Institute for Biobased Materials, Faculty of Science and Engineering, Maastricht University, Brightlands Chemelot Campus, 6167 RD, Geleen, The Netherlands

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Forckenbeckstraße 55, 52074, Aachen, Germany

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Aachen, Germany

3. Eva Miriam Buhl Aachen Institute of Pathology, Electron Microscopy Facility

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Institute of Pathology, Electron Microscopy Facility, RWTH Aachen University Hospital, Pauwelsstraße 30, 52074, Aachen, Germany

   Institute of Pathology, Electron Microscopy Facility, RWTH Aachen University Hospital, Aachen, Germany

4. Christian Gabriel Cornelissen Aachen Clinic for Pneumology and Internal Intensive Care Medicine (Medical Clinic V) Department of Biohybrid and Medical Textiles (BioTex) AME-Institute of Applied Medical Engineering

   Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Clinic for Pneumology and Internal Intensive Care Medicine (Medical Clinic V), RWTH Aachen University Hospital, Pauwelsstraße 30, 52074, Aachen, Germany

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Forckenbeckstraße 55, 52074, Aachen, Germany

   Clinic for Pneumology and Internal Intensive Care Medicine (Medical Clinic V), RWTH Aachen University Hospital, Aachen, Germany

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Aachen, Germany

5. Thomas Schmitz‐Rode Aachen Department of Biohybrid and Medical Textiles (BioTex) AME-Institute of Applied Medical Engineering

   RWTH Aachen University

   Affiliation as printed

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Forckenbeckstraße 55, 52074, Aachen, Germany

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Aachen, Germany

6. Stefan Jockenhoevel corresponding Aachen Department of Biohybrid and Medical Textiles (BioTex) AME-Institute of Applied Medical Engineering

   Maastricht University · RWTH Aachen University

   Affiliation as printed

   Aachen-Maastricht Institute for Biobased Materials, Faculty of Science and Engineering, Maastricht University, Brightlands Chemelot Campus, 6167 RD, Geleen, The Netherlands. jockenhoevel@ame.rwth-aachen.de

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Forckenbeckstraße 55, 52074, Aachen, Germany. jockenhoevel@ame.rwth-aachen.de

   Aachen-Maastricht Institute for Biobased Materials, Faculty of Science and Engineering, Maastricht University, Brightlands Chemelot Campus, 6167 RD, Geleen, The Netherlands

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Aachen, Germany

7. Anja Lena Thiebes Aachen Department of Biohybrid and Medical Textiles (BioTex) AME-Institute of Applied Medical Engineering

   Maastricht University · RWTH Aachen University

   Affiliation as printed

   Aachen-Maastricht Institute for Biobased Materials, Faculty of Science and Engineering, Maastricht University, Brightlands Chemelot Campus, 6167 RD, Geleen, The Netherlands

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Forckenbeckstraße 55, 52074, Aachen, Germany

   Department of Biohybrid and Medical Textiles (BioTex), AME - Institute of Applied Medical Engineering, Helmholtz Institute, RWTH Aachen University, Aachen, Germany