Malik Aydin1, Ella A Naumova2, Aliyah Bellm3, Ann-Kathrin Behrendt4, Federica Giachero5, Nora Bahlmann6, Wenli Zhang7, Stefan Wirth8, Anja Ehrhardt9, Wolfgang H Arnold10, Friedrich Paulsen11. 1. Laboratory of Experimental Pediatric Pneumology and Allergology, Center for Biomedical Education and Research, School of Life Sciences (ZBAF), Faculty of Health, Department of Human Medicine, Witten/Herdecke University, 42283 Wuppertal, Germany; Department of Pediatric Pneumology and Cardiology, Children's Hospital, Center for Clinical and Translational Research (CCTR), Helios University Hospital Wuppertal, 42283 Wuppertal, Germany. Electronic address: malik.aydin@uni-wh.de. 2. Department of Biological and Material Sciences in Dentistry, Faculty of Health, Witten/Herdecke University, 58455 Witten, Germany. Electronic address: ella.naumova@uni-wh.de. 3. Helios Hospital Krefeld, Children's Hospital, Teaching Hospital of RTWH University Hospital Aachen, 47805 Krefeld, Germany. Electronic address: aliyah.j.sanders@gmail.com. 4. Pediatric Rheumatology and Immunology, Department of Pediatrics, University Medicine Greifswald, 17475 Greifswald, Germany. Electronic address: ann-kathrin.behrendt@med.uni-greifswald.de. 5. Laboratory of Clinical Molecular Genetics and Epigenetics, Center for Biomedical Education and Research, School of Life Sciences (ZBAF), Faculty of Health, Witten/Herdecke University, 42283 Wuppertal, Germany. Electronic address: federica.giachero@uni-wh.de. 6. Institute of Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten/Herdecke University, 58453 Witten, Germany. Electronic address: nora.bahlmann@uni-wh.de. 7. Institute of Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten/Herdecke University, 58453 Witten, Germany. Electronic address: wenli.zhang@uni-wh.de. 8. Center for Child and Adolescent Medicine, Helios University Hospital Wuppertal, Witten/Herdecke University, 42283 Wuppertal, Germany. Electronic address: stefan.wirth@helios-gesundheit.de. 9. Institute of Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, Witten/Herdecke University, 58453 Witten, Germany. Electronic address: anja.ehrhardt@uni-wh.de. 10. Department of Biological and Material Sciences in Dentistry, Faculty of Health, Witten/Herdecke University, 58455 Witten, Germany. Electronic address: wolfgang.arnold@uni-wh.de. 11. Institute of Functional and Clinical Anatomy, Friedrich Alexander University Erlangen-Nurnberg, 91054 Erlangen, Germany; Sechenov University, Department of Topographic Anatomy and Operative Surgery, 119146 Moscow, Russia. Electronic address: friedrich.paulsen@fau.de.
Abstract
OBJECTIVES: Research involving the nose reveals important information regarding the morphology and physiology of the epithelium and its molecular response to agents. The role of nasal epithelial cells and other cell subsets within the nasal epithelium play an interesting translational split between experimental and clinical research studying respiratory disorders or pathogen reactions. With an additional technical manuscript including a detailed description of important technical aspects, tips, tricks, and nuances for a successful culturing of primary, human nasal epithelial cells (NAEPCs), we here aim to improve the process of communication between experimentalists and physicians, supporting the purpose of a fruitful work for future translational projects. METHODS: Based on previous work on various complex culture models of subject-derived NAEPCs, this additional manuscript harmonizes previously published facts combined with own experiences for a trouble-free implementation in laboratories. RESULTS: A well-designed experimental question is essential prior to the establishment of different NAEPCs culture models. The correct method of cell extraction from the nasal cavity is essential and represent an important basis for successful culture work. Prior enzymatic processing of biopsy specimens, cell culture materials, collagenization procedure, culture conditions, and choice of culture medium are some important practical notes that increase the quality of the culture. Moreover, protocols on imaging techniques including histologic and electron microscopy must be adapted for NAEPC culture. Adapted flow cytometric protocols and transepithelial electrical resistance measurements can add valuable information. OUTLOOK: A successful culturing of NAEPCs can provide an important basis for genetic studies and the implementation of omics-science, which is increasingly receiving broad attention in the scientific community. The common aim of in vitro 'mini-noses' will be a breakthrough in laboratories aiming to perform research under in vivo conditions. Here, organoid models are interesting models presenting a basis for translational studies.
OBJECTIVES: Research involving the nose reveals important information regarding the morphology and physiology of the epithelium and its molecular response to agents. The role of nasal epithelial cells and other cell subsets within the nasal epithelium play an interesting translational split between experimental and clinical research studying respiratory disorders or pathogen reactions. With an additional technical manuscript including a detailed description of important technical aspects, tips, tricks, and nuances for a successful culturing of primary, human nasal epithelial cells (NAEPCs), we here aim to improve the process of communication between experimentalists and physicians, supporting the purpose of a fruitful work for future translational projects. METHODS: Based on previous work on various complex culture models of subject-derived NAEPCs, this additional manuscript harmonizes previously published facts combined with own experiences for a trouble-free implementation in laboratories. RESULTS: A well-designed experimental question is essential prior to the establishment of different NAEPCs culture models. The correct method of cell extraction from the nasal cavity is essential and represent an important basis for successful culture work. Prior enzymatic processing of biopsy specimens, cell culture materials, collagenization procedure, culture conditions, and choice of culture medium are some important practical notes that increase the quality of the culture. Moreover, protocols on imaging techniques including histologic and electron microscopy must be adapted for NAEPC culture. Adapted flow cytometric protocols and transepithelial electrical resistance measurements can add valuable information. OUTLOOK: A successful culturing of NAEPCs can provide an important basis for genetic studies and the implementation of omics-science, which is increasingly receiving broad attention in the scientific community. The common aim of in vitro 'mini-noses' will be a breakthrough in laboratories aiming to perform research under in vivo conditions. Here, organoid models are interesting models presenting a basis for translational studies.
Authors: Malik Aydin; Jana Dietrich; Joana Witt; Maximiliane S C Finkbeiner; Jonas J-H Park; Stefan Wirth; Christine E Engeland; Friedrich Paulsen; Anja Ehrhardt Journal: Int J Mol Sci Date: 2021-11-30 Impact factor: 5.923