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Tytuł:
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An optimized, robust and reproducible protocol to generate well-differentiated primary nasal epithelial models from extremely premature infants.
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Autorzy:
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Martens A; Division of Neonatology, Pediatric Intensive Care & Neuropediatrics, Comprehensive Center for Pediatrics, Medical University of Vienna, Vienna, Austria.
Amann G; Institute of Clinical Pathology, Medical University of Vienna, Vienna, Austria.
Schmidt K; Center for Anatomy and Cell Biology, Medical University of Vienna, Vienna, Austria.
Gaupmann R; Division of Pediatric Pulmonology, Allergology and Endocrinology, Comprehensive Center for Pediatrics, Medical University of Vienna, Vienna, Austria.
Böhm B; Division of Pediatric Pulmonology, Allergology and Endocrinology, Comprehensive Center for Pediatrics, Medical University of Vienna, Vienna, Austria.
Dehlink E; Division of Pediatric Pulmonology, Allergology and Endocrinology, Comprehensive Center for Pediatrics, Medical University of Vienna, Vienna, Austria.
Szépfalusi Z; Division of Pediatric Pulmonology, Allergology and Endocrinology, Comprehensive Center for Pediatrics, Medical University of Vienna, Vienna, Austria.
Förster-Waldl E; Division of Neonatology, Pediatric Intensive Care & Neuropediatrics, Comprehensive Center for Pediatrics, Medical University of Vienna, Vienna, Austria.; Center for Congenital Immunodeficiencies, Medical University of Vienna, Vienna, Austria.
Berger A; Division of Neonatology, Pediatric Intensive Care & Neuropediatrics, Comprehensive Center for Pediatrics, Medical University of Vienna, Vienna, Austria.
Fyhrquist N; Department of Bacteriology and Immunology, University of Helsinki, Helsinki, Finland.; Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
Alenius H; Department of Bacteriology and Immunology, University of Helsinki, Helsinki, Finland.; Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.
Wisgrill L; Division of Neonatology, Pediatric Intensive Care & Neuropediatrics, Comprehensive Center for Pediatrics, Medical University of Vienna, Vienna, Austria. .
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Źródło:
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Scientific reports [Sci Rep] 2019 Dec 27; Vol. 9 (1), pp. 20069. Date of Electronic Publication: 2019 Dec 27.
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Typ publikacji:
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Journal Article; Research Support, Non-U.S. Gov't
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Język:
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English
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Imprint Name(s):
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Original Publication: London : Nature Publishing Group, copyright 2011-
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MeSH Terms:
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Cell Differentiation*
Models, Biological*
Nasal Mucosa/*cytology
Adult ; Case-Control Studies ; Cells, Cultured ; Disease Susceptibility ; Humans ; Infant, Extremely Premature ; Infant, Newborn ; Reproducibility of Results ; Respiratory Tract Diseases/etiology ; Respiratory Tract Diseases/immunology
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References:
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Entry Date(s):
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Date Created: 20191229 Date Completed: 20201207 Latest Revision: 20210110
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Update Code:
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20240104
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PubMed Central ID:
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PMC6934534
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DOI:
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10.1038/s41598-019-56737-9
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PMID:
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31882915
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Extremely premature infants are prone to severe respiratory infections, and the mechanisms underlying this exceptional susceptibility are largely unknown. Nasal epithelial cells (NEC) represent the first-line of defense and adult-derived ALI cell culture models show promising results in mimicking in vivo physiology. Therefore, the aim of this study was to develop a robust and reliable protocol for generating well-differentiated cell culture models from NECs of extremely premature infants. Nasal brushing was performed in 13 extremely premature infants at term corrected age and in 11 healthy adult controls to obtain NECs for differentiation at air-liquid interface (ALI). Differentiation was verified using imaging and functional analysis. Successful isolation and differentiation was achieved for 5 (38.5%) preterm and 5 (45.5%) adult samples. Preterm and adult ALI-cultures both showed well-differentiated morphology and ciliary function, however, preterm cultures required significantly longer cultivation times for acquiring full differentiation (44 ± 3.92 vs. 23 ± 1.83 days; p < 0.0001). Moreover, we observed that recent respiratory support may impair successful NECs isolation. Herewithin, we describe a safe, reliable and reproducible method to generate well-differentiated ALI-models from NECs of extremely premature infants. These models provide a valuable foundation for further studies regarding immunological and inflammatory responses and respiratory disorders in extremely premature infants.
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