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  1. Article ; Online: Temporal Whole-Transcriptomic Analysis of Characterized In Vitro and Ex Vivo Primary Nasal Epithelia

    Jelmer Legebeke / Katie L. Horton / Claire L. Jackson / Janice Coles / Amanda Harris / Htoo A. Wai / John W. Holloway / Gabrielle Wheway / Diana Baralle / Jane S. Lucas

    Frontiers in Cell and Developmental Biology, Vol

    2022  Volume 10

    Abstract: Air-liquid interface (ALI) cell culture of primary airway progenitors enables the differentiation and recapitulation of a pseudostratified epithelium in vitro, providing a highly useful tool for researching respiratory health and disease. Previous ... ...

    Abstract Air-liquid interface (ALI) cell culture of primary airway progenitors enables the differentiation and recapitulation of a pseudostratified epithelium in vitro, providing a highly useful tool for researching respiratory health and disease. Previous studies into gene expression in ALI-cultures compared to ex vivo nasal brushings have been limited in the number of time-points and/or the number of genes studied. In this study physiological and global transcriptomic changes were assessed in an extended in vitro 63-day human healthy nasal epithelium ALI-culture period and compared to ex vivo nasal brushing samples. Ex vivo nasal brushing samples formed distinct transcriptome clusters to in vitro ALI-cultured nasal epithelia, with from day 14 onwards ALI samples best matching the ex vivo samples. Immune response regulation genes were not expressed in the in vitro ALI-culture compared to the ex vivo nasal brushing samples, likely because the in vitro cultures lack an airway microbiome, lack airborne particles stimulation, or did not host an immune cell component. This highlights the need for more advanced co-cultures with immune cell representation to better reflect the physiological state. During the first week of ALI-culture genes related to metabolism and proliferation were increased. By the end of week 1 epithelial cell barrier function plateaued and multiciliated cell differentiation started, although widespread ciliation was not complete until day 28. These results highlight that time-points at which ALI-cultures are harvested for research studies needs to be carefully considered to suit the purpose of investigation (transcriptomic and/or functional analysis).
    Keywords primary nasal epithelium ; air-liquid interface culture ; airway cilia ; physiological analysis ; whole transcriptome analysis ; Biology (General) ; QH301-705.5
    Subject code 570
    Language English
    Publishing date 2022-06-01T00:00:00Z
    Publisher Frontiers Media S.A.
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: ERS International Congress 2021

    Oliver W. Meldrum / Kylie B.R. Belchamber / Kiarina D. Chichirelo-Konstantynovych / Katie L. Horton / Tetyana V. Konstantynovych / Merete B. Long / Melissa J. McDonnell / Lidia Perea / Alberto L. Garcia-Basteiro / Michael R. Loebinger / Raquel Duarte / Holly R. Keir

    ERJ Open Research, Vol 8, Iss

    highlights from the Respiratory Infections Assembly

    2022  Volume 2

    Abstract: The European Respiratory Society International Congress 2021 took place virtually for the second year running due to the coronavirus pandemic. The Congress programme featured more than 400 sessions and 3000 abstract presentations, covering the entire ... ...

    Abstract The European Respiratory Society International Congress 2021 took place virtually for the second year running due to the coronavirus pandemic. The Congress programme featured more than 400 sessions and 3000 abstract presentations, covering the entire field of respiratory science and medicine. In this article, early career members of the Respiratory Infections Assembly summarise a selection of sessions across a broad range of topics, including presentations on bronchiectasis, non-tuberculosis mycobacteria, tuberculosis, cystic fibrosis and COVID-19.
    Keywords Medicine ; R
    Language English
    Publishing date 2022-05-01T00:00:00Z
    Publisher European Respiratory Society
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: A Revised Protocol for Culture of Airway Epithelial Cells as a Diagnostic Tool for Primary Ciliary Dyskinesia

    Janice L. Coles / James Thompson / Katie L. Horton / Robert A. Hirst / Paul Griffin / Gwyneth M. Williams / Patricia Goggin / Regan Doherty / Peter M. Lackie / Amanda Harris / Woolf T. Walker / Christopher O’Callaghan / Claire Hogg / Jane S. Lucas / Cornelia Blume / Claire L. Jackson

    Journal of Clinical Medicine, Vol 9, Iss 3753, p

    2020  Volume 3753

    Abstract: Air–liquid interface (ALI) culture of nasal epithelial cells is a valuable tool in the diagnosis and research of primary ciliary dyskinesia (PCD). Ex vivo samples often display secondary dyskinesia from cell damage during sampling, infection or ... ...

    Abstract Air–liquid interface (ALI) culture of nasal epithelial cells is a valuable tool in the diagnosis and research of primary ciliary dyskinesia (PCD). Ex vivo samples often display secondary dyskinesia from cell damage during sampling, infection or inflammation confounding PCD diagnostic results. ALI culture enables regeneration of healthy cilia facilitating differentiation of primary from secondary ciliary dyskinesia. We describe a revised ALI culture method adopted from April 2018 across three collaborating PCD diagnostic sites, including current University Hospital Southampton COVID-19 risk mitigation measures, and present results. Two hundred and forty nasal epithelial cell samples were seeded for ALI culture and 199 (82.9%) were ciliated. Fifty-four of 83 (63.9%) ex vivo samples which were originally equivocal or insufficient provided diagnostic information following in vitro culture. Surplus basal epithelial cells from 181 nasal brushing samples were frozen in liquid nitrogen; 39 samples were ALI-cultured after cryostorage and all ciliated. The ciliary beat patterns of ex vivo samples (by high-speed video microscopy) were recapitulated, scanning electron microscopy demonstrated excellent ciliation, and cilia could be immuno-fluorescently labelled (anti-alpha-tubulin and anti-RSPH4a) in representative cases that were ALI-cultured after cryostorage. In summary, our ALI culture protocol provides high ciliation rates across three centres, minimising patient recall for repeat brushing biopsies and improving diagnostic certainty. Cryostorage of surplus diagnostic samples was successful, facilitating PCD research.
    Keywords PCD ; ALI culture ; bio-resource ; primary nasal epithelium ; diagnostics ; Medicine ; R
    Subject code 610
    Language English
    Publishing date 2020-11-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: Proceedings of the 4th BEAT-PCD Conference and 5th PCD Training School

    Laura E. Gardner / Katie L. Horton / Amelia Shoemark / Jane S. Lucas / Kim G. Nielsen / Helene Kobbernagel / Bruna Rubbo / Robert A. Hirst / Panayiotis Kouis / Nicola Ullmann / Ana Reula / Nisreen Rumman / Hannah M. Mitchison / Andreia Pinto / Charlotte Richardson / Anne Schmidt / James Thompson / René Gaupmann / Maciej Dabrowski /
    Pleasantine Mill / Siobhan B. Carr / Dominic P. Norris / Claudia E. Kuehni / Myrofora Goutaki / Claire Hogg

    BMC Proceedings, Vol 14, Iss S8, Pp 1-

    2020  Volume 17

    Abstract: Abstract Primary ciliary dyskinesia (PCD) is an inherited ciliopathy leading to chronic suppurative lung disease, chronic rhinosinusitis, middle ear disease, sub-fertility and situs abnormalities. As PCD is rare, it is important that scientists and ... ...

    Abstract Abstract Primary ciliary dyskinesia (PCD) is an inherited ciliopathy leading to chronic suppurative lung disease, chronic rhinosinusitis, middle ear disease, sub-fertility and situs abnormalities. As PCD is rare, it is important that scientists and clinicians foster international collaborations to share expertise in order to provide the best possible diagnostic and management strategies. ‘Better Experimental Approaches to Treat Primary Ciliary Dyskinesia’ (BEAT-PCD) is a multidisciplinary network funded by EU COST Action (BM1407) to coordinate innovative basic science and clinical research from across the world to drive advances in the field. The fourth and final BEAT-PCD Conference and fifth PCD Training School were held jointly in March 2019 in Poznan, Poland. The varied program of plenaries, workshops, break-out sessions, oral and poster presentations were aimed to enhance the knowledge and skills of delegates, whilst also providing a collaborative platform to exchange ideas. In this final BEAT-PCD conference we were able to build upon programmes developed throughout the lifetime of the COST Action. These proceedings report on the conference, highlighting some of the successes of the BEAT-PCD programme.
    Keywords Primary ciliary dyskinesia ; Chronic respiratory disease ; Multidisciplinary ; Medicine ; R ; Science ; Q
    Language English
    Publishing date 2020-06-01T00:00:00Z
    Publisher BMC
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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