title: Temporal Monitoring of Differentiated Human Airway Epithelial Cells Using Microfluidics.
abstract: The airway epithelium is exposed to a variety of harmful agents during breathing and appropriate cellular responses are essential to maintain tissue homeostasis. Recent evidence has highlighted the contribution of epithelial barrier dysfunction in the development of many chronic respiratory diseases. Despite intense research efforts, the responses of the airway barrier to environmental agents are not fully understood, mainly due to lack of suitable in vitro models that recapitulate the complex in vivo situation accurately. Using an interdisciplinary approach, we describe a novel dynamic 3D in vitro model of the airway epithelium, incorporating fully differentiated primary human airway epithelial cells at the air-liquid interface and a basolateral microfluidic supply of nutrients simulating the interstitial flow observed in vivo. Through combination of the microfluidic culture system with an automated fraction collector the kinetics of cellular responses by the airway epithelium to environmental agents can be analysed at the early phases for the first time and with much higher sensitivity compared to common static in vitro models. Following exposure of primary differentiated epithelial cells to pollen we show that CXCL8/IL-8 release is detectable within the first 2h and peaks at 4-6h under microfluidic conditions, a response which was not observed in conventional static culture conditions. Such a microfluidic culture model is likely to have utility for high resolution temporal profiling of toxicological and pharmacological responses of the airway epithelial barrier, as well as for studies of disease mechanisms.
authors.affiliation: Academic Unit of Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton, United Kingdom.
authors.forename: Cornelia
authors.initials: C
authors.lastname: Blume
authors.affiliation: Electronics and Computer Sciences, Faculty of Physical Sciences and Engineering, University of Southampton, Southampton, United Kingdom.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Reale
authors.affiliation: Electronics and Computer Sciences, Faculty of Physical Sciences and Engineering, University of Southampton, Southampton, United Kingdom.
authors.forename: Marie
authors.initials: M
authors.lastname: Held
authors.affiliation: Academic Unit of Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton, United Kingdom.
authors.forename: Timothy M
authors.initials: TM
authors.lastname: Millar
authors.affiliation: Academic Unit of Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton, United Kingdom.
authors.forename: Jane E
authors.initials: JE
authors.lastname: Collins
authors.affiliation: Academic Unit of Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton, United Kingdom; Institute for Life Sciences, University of Southampton, Southampton, United Kingdom; National Institute for Health Research, Respiratory Biomedical Research Unit, University Hospital Southampton, Southampton, United Kingdom.
authors.forename: Donna E
authors.initials: DE
authors.lastname: Davies
authors.affiliation: Electronics and Computer Sciences, Faculty of Physical Sciences and Engineering, University of Southampton, Southampton, United Kingdom; Institute for Life Sciences, University of Southampton, Southampton, United Kingdom.
authors.forename: Hywel
authors.initials: H
authors.lastname: Morgan
authors.affiliation: Academic Unit of Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, Southampton, United Kingdom; Institute for Life Sciences, University of Southampton, Southampton, United Kingdom; National Institute for Health Research, Respiratory Biomedical Research Unit, University Hospital Southampton, Southampton, United Kingdom.
authors.forename: Emily J
authors.initials: EJ
authors.lastname: Swindle
mesh_terms: D004847:Epithelial Cells / Q000166:cytology*; D006801:Humans; D066298:In Vitro Techniques; D044085:Microfluidics; D012137:Respiratory System / Q000166:cytology*