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

David Miller, Steve W Turner, Daniella Spiteri Cornish, Neil McInnes, Alison Scaife, Peter J Danielian, Graham Devereux, Garry M Walsh

Research output: Contribution to journalArticle

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Abstract

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-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.

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 “naïve” cells that may prove useful in elucidating the role of the epithelium in the early origins of asthma and/or allergic rhinitis.
Original languageEnglish
Article numbere78321
Number of pages7
JournalPloS ONE
Volume8
Issue number11
DOIs
Publication statusPublished - 4 Nov 2013

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neonates
epithelial cells
Epithelial Cells
Parturition
Newborn Infant
dust mites
interleukin-8
interleukin-1
Pyroglyphidae
Interleukin-8
Interleukin-1
Dust
Nose
Pulmonary diseases
rhinitis
extracts
asthma
immunocytochemistry
cells
Brushes

Cite this

Culture of airway epithelial cells from neonates sampled within 48-hours of birth. / Miller, David; Turner, Steve W; Spiteri Cornish, Daniella; McInnes, Neil; Scaife, Alison; Danielian, Peter J ; Devereux, Graham; Walsh, Garry M.

In: PloS ONE, Vol. 8, No. 11, e78321, 04.11.2013.

Research output: Contribution to journalArticle

Miller D, Turner SW, Spiteri Cornish D, McInnes N, Scaife A, Danielian PJ et al. Culture of airway epithelial cells from neonates sampled within 48-hours of birth. PloS ONE. 2013 Nov 4;8(11). e78321. https://doi.org/10.1371/journal.pone.0078321
Miller, David ; Turner, Steve W ; Spiteri Cornish, Daniella ; McInnes, Neil ; Scaife, Alison ; Danielian, Peter J ; Devereux, Graham ; Walsh, Garry M. / Culture of airway epithelial cells from neonates sampled within 48-hours of birth. In: PloS ONE. 2013 ; Vol. 8, No. 11.
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