Culture of airway epithelial cells from neonates sampled within 48-hours of birth

• Published in: PloS one 2013-11, Vol.8 (11), p.e78321-e78321
• Main Authors: Miller, David, Turner, Steve W, Spiteri-Cornish, Daniella, McInnes, Neil, Scaife, Alison, Danielian, Peter J, Devereux, Graham, Walsh, Garry M
• Format: Article
• Language: English
• Subjects: Allergic rhinitis; Allergies; Animals; Asthma; Cell culture; Cell Proliferation; Children & youth; Childrens health; Cohort analysis; Complex Mixtures - chemistry; Complex Mixtures - pharmacology; Confluence; Consent; Dose-Response Relationship, Immunologic; Dust; Electron microscopy; Epithelial cells; Epithelial Cells - cytology; Epithelial Cells - drug effects; Epithelial Cells - immunology; Epithelium; Female; Health risk assessment; Hospitals; House dust; Humans; Immunocytochemistry; Infant, Newborn; Infants; Inflammation; Interleukin 8; Interleukin-1beta - pharmacology; Interleukin-8 - biosynthesis; Interleukin-8 - metabolism; Male; Medical research; Monomolecular films; Nasal Cavity - cytology; Nasal Cavity - immunology; Neonates; Newborn babies; Newborn infants; Nose; Pathogenesis; Pediatrics; Phenotypes; Primary Cell Culture; Pyroglyphidae - chemistry; Respiratory Mucosa - cytology; Respiratory Mucosa - drug effects; Respiratory Mucosa - immunology; Respiratory tract; Rhinitis; Science; Smoking; Tumor necrosis factor; Tumor Necrosis Factor-alpha - pharmacology; Tumor necrosis factor-α; Womens health
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0078321

Little is known about how neonatal airway epithelial cell phenotype impacts on respiratory disease in later life. This study aimed to establish a methodology to culture and characterise neonatal nasal epithelial cells sampled from healthy, non-sedated infants within 48 hours of delivery. Nasal epithelial cells were sampled by brushing both nostrils with an interdental brush, grown to confluence and sub-cultured. Cultured cells were characterised morphologically by light and electron microscopy and by immunocytochemistry. As an exemplar pro-inflammatory chemokine, IL-8 concentrations were measured in supernatants from unstimulated monolayers and after exposure to IL-1β/TNF-α or house dust mite extract. Primary cultures were successfully established in 135 (91%) of 149 neonatal samples seeded, with 79% (n  =  117) successfully cultured to passage 3. The epithelial lineage of the cells was confirmed by morphological analysis and immunostaining. Constitutive IL-8 secretion was observed and was upregulated by IL-1β/TNF-α or house dust mite extract in a dose dependent manner. We describe a safe, minimally invasive method of culturing nasal epithelial cells from neonates suitable for functional cell analysis offering an opportunity to study "naïve" cells that may prove useful in elucidating the role of the epithelium in the early origins of asthma and/or allergic rhinitis.