Culture of airway epithelial cells from neonates sampled within 48-hours of birth.
BACKGROUND
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-1β/TNF-α 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-1β/TNF-α or house dust mite extract in a dose dependent manner.


CONCLUSIONS
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.
Department of Child Health, Royal Aberdeen Children's Hospital, University of Aberdeen, Aberdeen, United Kingdom ; Institute of Medical Science, University of Aberdeen, Aberdeen, United Kingdom.
David
D
Miller
Steve W
SW
Turner
Daniella
D
Spiteri-Cornish
Neil
N
McInnes
Alison
A
Scaife
Peter J
PJ
Danielian
Graham
G
Devereux
Garry M
GM
Walsh
D000818:Animals; D049109:Cell Proliferation; D045424:Complex Mixtures / Q000737:chemistry / Q000494:pharmacology*; D004306:Dose-Response Relationship, Immunologic; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D005260:Female; D006801:Humans; D007231:Infant, Newborn; D053583:Interleukin-1beta / Q000494:pharmacology*; D016209:Interleukin-8 / Q000096:biosynthesis / Q000378:metabolism; D008297:Male; D009296:Nasal Cavity / Q000166:cytology / Q000276:immunology; D061251:Primary Cell Culture; D039981:Pyroglyphidae / Q000737:chemistry; D020545:Respiratory Mucosa / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D045424:Complex Mixtures; D053583:Interleukin-1beta; D016209:Interleukin-8; D014409:Tumor Necrosis Factor-alpha