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Clonally expanded human airway smooth muscle cells exhibit morphological and functional heterogeneity

BACKGROUND: Mesenchyme-derived airway cell populations including airway smooth muscle (ASM) cells, fibroblasts and myofibroblasts play key roles in the pathogenesis of airway inflammation and remodeling. Phenotypic and functional characterisation of these cell populations are confounded by their het...

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Autores principales: Singh, Shailendra R, Billington, Charlotte K, Sayers, Ian, Hall, Ian P
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4014754/
https://www.ncbi.nlm.nih.gov/pubmed/24886333
http://dx.doi.org/10.1186/1465-9921-15-57
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author Singh, Shailendra R
Billington, Charlotte K
Sayers, Ian
Hall, Ian P
author_facet Singh, Shailendra R
Billington, Charlotte K
Sayers, Ian
Hall, Ian P
author_sort Singh, Shailendra R
collection PubMed
description BACKGROUND: Mesenchyme-derived airway cell populations including airway smooth muscle (ASM) cells, fibroblasts and myofibroblasts play key roles in the pathogenesis of airway inflammation and remodeling. Phenotypic and functional characterisation of these cell populations are confounded by their heterogeneity in vitro. It is unclear which mechanisms underlie the creation of these different sub-populations. The study objectives were to investigate whether ASM cells are capable of clonal expansion and if so (i) what proportion possess this capability and (ii) do clonal populations exhibit variation in terms of morphology, phenotype, proliferation rates and pro-relaxant or pro-contractile signaling pathways. METHODS: Early passage human ASM cells were subjected to single-cell cloning and their doubling time was recorded. Immunocytochemistry was performed to assess localization and levels of markers previously reported to be specifically associated with smooth muscle or fibroblasts. Finally functional assays were used to reveal differences between clonal populations specifically assessing mitogen-induced proliferation and pro-relaxant and pro-contractile signaling pathways. RESULTS: Our studies provide evidence that a high proportion (58%) of single cells present within early passage human ASM cell cultures have the potential to create expanded cell populations. Despite being clonally-originated, morphological heterogeneity was still evident within these clonal populations as assessed by the range in expression of markers associated with smooth muscle cells. Functional diversity was observed between clonal populations with 10 μM isoproterenol-induced cyclic AMP responses ranging from 1.4 - 5.4 fold cf basal and bradykinin-induced inositol phosphate from 1.8 - 5.2 fold cf basal. CONCLUSION: In summary we show for the first time that primary human ASM cells are capable of clonal expansion and that the resulting clonal populations themselves exhibit phenotypic plasticity.
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spelling pubmed-40147542014-05-10 Clonally expanded human airway smooth muscle cells exhibit morphological and functional heterogeneity Singh, Shailendra R Billington, Charlotte K Sayers, Ian Hall, Ian P Respir Res Research BACKGROUND: Mesenchyme-derived airway cell populations including airway smooth muscle (ASM) cells, fibroblasts and myofibroblasts play key roles in the pathogenesis of airway inflammation and remodeling. Phenotypic and functional characterisation of these cell populations are confounded by their heterogeneity in vitro. It is unclear which mechanisms underlie the creation of these different sub-populations. The study objectives were to investigate whether ASM cells are capable of clonal expansion and if so (i) what proportion possess this capability and (ii) do clonal populations exhibit variation in terms of morphology, phenotype, proliferation rates and pro-relaxant or pro-contractile signaling pathways. METHODS: Early passage human ASM cells were subjected to single-cell cloning and their doubling time was recorded. Immunocytochemistry was performed to assess localization and levels of markers previously reported to be specifically associated with smooth muscle or fibroblasts. Finally functional assays were used to reveal differences between clonal populations specifically assessing mitogen-induced proliferation and pro-relaxant and pro-contractile signaling pathways. RESULTS: Our studies provide evidence that a high proportion (58%) of single cells present within early passage human ASM cell cultures have the potential to create expanded cell populations. Despite being clonally-originated, morphological heterogeneity was still evident within these clonal populations as assessed by the range in expression of markers associated with smooth muscle cells. Functional diversity was observed between clonal populations with 10 μM isoproterenol-induced cyclic AMP responses ranging from 1.4 - 5.4 fold cf basal and bradykinin-induced inositol phosphate from 1.8 - 5.2 fold cf basal. CONCLUSION: In summary we show for the first time that primary human ASM cells are capable of clonal expansion and that the resulting clonal populations themselves exhibit phenotypic plasticity. BioMed Central 2014 2014-05-03 /pmc/articles/PMC4014754/ /pubmed/24886333 http://dx.doi.org/10.1186/1465-9921-15-57 Text en Copyright © 2014 Singh et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Singh, Shailendra R
Billington, Charlotte K
Sayers, Ian
Hall, Ian P
Clonally expanded human airway smooth muscle cells exhibit morphological and functional heterogeneity
title Clonally expanded human airway smooth muscle cells exhibit morphological and functional heterogeneity
title_full Clonally expanded human airway smooth muscle cells exhibit morphological and functional heterogeneity
title_fullStr Clonally expanded human airway smooth muscle cells exhibit morphological and functional heterogeneity
title_full_unstemmed Clonally expanded human airway smooth muscle cells exhibit morphological and functional heterogeneity
title_short Clonally expanded human airway smooth muscle cells exhibit morphological and functional heterogeneity
title_sort clonally expanded human airway smooth muscle cells exhibit morphological and functional heterogeneity
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4014754/
https://www.ncbi.nlm.nih.gov/pubmed/24886333
http://dx.doi.org/10.1186/1465-9921-15-57
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