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

Abstract

IntroductionLittle 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.MethodsNasal 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.ResultsPrimary 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.ConclusionWe 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.

Characterisation of morphological differences in well-differentiated nasal epithelial cell cultures from preterm and term infants at birth and one-year

“…Nasal sampling was performed either with the neonate lying in a parent’s arms or in a cot using the technique described by Miller et al . (7) In brief, the infant’s head was gently secured using one hand and a 2.7 mm diameter interdental brush (DentoCare Professional, London, UK) was introduced into each nostril (one brush/nostril) in turn and gently rotated twice against the medial aspect of the inferior turbinate.…”

“…Recent publications have highlighted the potential benefit of this technique in investigating early changes in immune function in cystic fibrosis (CF) infants, in the study of the nasal transcriptome of infants, and in the innate immune responses of the airway epithelium to allergens as part of asthma pathogenesis research. (7,9,10) Mosler et al described the culture of nasal epithelial cells from CF infants, some of whom were as young as one-month old, but to date, Miller et al is the only publication describing monolayer culture of nasal epithelial cells from newborn infants. (7)…”

“…(7,9,10) Mosler et al described the culture of nasal epithelial cells from CF infants, some of whom were as young as one-month old, but to date, Miller et al is the only publication describing monolayer culture of nasal epithelial cells from newborn infants. (7)…”

Pro‐inflammatory mediator responses from neonatal airway epithelial cells and early childhood wheeze

“…Recruitment took place at 10‐12 weeks gestation, and details of parental asthma, smoking and socioeconomic status were collected. Within 48 h of birth, nasal AEC were obtained by brushing and cultured in a submerged model . In a subset of individuals, there were sufficient cells available for cultured AEC to also be exposed to lipopolysaccharide (LPS).…”

“…Within 48 h of birth, nasal AEC were obtained by brushing and cultured in a submerged model. 16 In a subset of individuals, there were sufficient cells available for cultured AEC to also be exposed to lipopolysaccharide (LPS). Close to the child's fourth birthday a postal questionnaire was completed.…”