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

Singh, Shailendra R.; Billington, Charlotte K.; Sayers, Ian; Hall, Ian P.

Authors

Shailendra R. Singh

Charlotte K. Billington

Ian P. Hall



Abstract

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.

Journal Article Type Article
Publication Date May 3, 2014
Journal Respiratory Research
Print ISSN 1465-9921
Electronic ISSN 1465-993X
Publisher Humana Press
Peer Reviewed Peer Reviewed
Volume 15
Issue 1
Article Number 57
APA6 Citation Singh, S. R., Billington, C. K., Sayers, I., & Hall, I. P. (2014). Clonally expanded human airway smooth muscle cells exhibit morphological and functional heterogeneity. Respiratory Research, 15(1), doi:10.1186/1465-9921-15-57
DOI https://doi.org/10.1186/1465-9921-15-57
Keywords Human airway smooth muscle, Clonal expansion, Phenotype, Plasticity, Remodleing
Publisher URL http://dx.doi.org/10.1186/1465-9921-15-57
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0





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