Culture of Airway Epithelial Cells from Neonates Sampled within 48-Hours of Birth

Introduction: 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. Methods: 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-1b/TNF-a or house dust mite extract. Results: 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-1b/TNF-a or house dust mite extract in a dose dependent manner. Conclusion: We describe a safe, minimally invasive method of culturing nasal epithelial cells from neonates suitable for functional cell analysis offering an opportunity to study ‘‘nao ¨ve’’ cells that may prove useful in elucidating the role of the epithelium in the early origins of asthma and/or allergic rhinitis.