Immortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro

Abstract To overcome the scarcity of primary human alveolar epithelial cells for lung research, and the limitations of current cell lines to recapitulate the phenotype, functional and molecular characteristics of the healthy human alveolar epithelium, we have developed a new method to immortalise pr...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Alberto Katsumiti, Pakatip Ruenraroengsak, Miren P. Cajaraville, Andrew J. Thorley, Teresa D. Tetley
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/937c8d854bab44f99579b0458b77380f
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:937c8d854bab44f99579b0458b77380f
record_format	dspace
spelling	oai:doaj.org-article:937c8d854bab44f99579b0458b77380f2021-12-02T11:40:41ZImmortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro10.1038/s41598-020-77191-y2045-2322https://doaj.org/article/937c8d854bab44f99579b0458b77380f2020-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-77191-yhttps://doaj.org/toc/2045-2322Abstract To overcome the scarcity of primary human alveolar epithelial cells for lung research, and the limitations of current cell lines to recapitulate the phenotype, functional and molecular characteristics of the healthy human alveolar epithelium, we have developed a new method to immortalise primary human alveolar epithelial lung cells using a non-viral vector to transfect the telomerase catalytic subunit (hTERT) and the simian virus 40 large-tumour antigen (SV40). Twelve strains of immortalised cells (ICs) were generated and characterised using molecular, immunochemical and morphological techniques. Cell proliferation and sensitivity to polystyrene nanoparticles (PS) were evaluated. ICs expressed caveolin-1, podoplanin and receptor for advanced glycation end-products (RAGE), and most cells were negative for alkaline phosphatase staining, indicating characteristics of AT1-like cells. However, most strains also contained some cells that expressed pro-surfactant protein C, classically described to be expressed only by AT2 cells. Thus, the ICs mimic the cellular heterogeneity in the human alveolar epithelium. These ICs can be passaged, replicate rapidly and remain confluent beyond 15 days. ICs showed differential sensitivity to positive and negatively charged PS nanoparticles, illustrating their potential value as an in vitro model to study respiratory bioreactivity. These novel ICs offer a unique resource to study human alveolar epithelial biology.Alberto KatsumitiPakatip RuenraroengsakMiren P. CajaravilleAndrew J. ThorleyTeresa D. TetleyNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-13 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Alberto Katsumiti Pakatip Ruenraroengsak Miren P. Cajaraville Andrew J. Thorley Teresa D. Tetley Immortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro
description	Abstract To overcome the scarcity of primary human alveolar epithelial cells for lung research, and the limitations of current cell lines to recapitulate the phenotype, functional and molecular characteristics of the healthy human alveolar epithelium, we have developed a new method to immortalise primary human alveolar epithelial lung cells using a non-viral vector to transfect the telomerase catalytic subunit (hTERT) and the simian virus 40 large-tumour antigen (SV40). Twelve strains of immortalised cells (ICs) were generated and characterised using molecular, immunochemical and morphological techniques. Cell proliferation and sensitivity to polystyrene nanoparticles (PS) were evaluated. ICs expressed caveolin-1, podoplanin and receptor for advanced glycation end-products (RAGE), and most cells were negative for alkaline phosphatase staining, indicating characteristics of AT1-like cells. However, most strains also contained some cells that expressed pro-surfactant protein C, classically described to be expressed only by AT2 cells. Thus, the ICs mimic the cellular heterogeneity in the human alveolar epithelium. These ICs can be passaged, replicate rapidly and remain confluent beyond 15 days. ICs showed differential sensitivity to positive and negatively charged PS nanoparticles, illustrating their potential value as an in vitro model to study respiratory bioreactivity. These novel ICs offer a unique resource to study human alveolar epithelial biology.
format	article
author	Alberto Katsumiti Pakatip Ruenraroengsak Miren P. Cajaraville Andrew J. Thorley Teresa D. Tetley
author_facet	Alberto Katsumiti Pakatip Ruenraroengsak Miren P. Cajaraville Andrew J. Thorley Teresa D. Tetley
author_sort	Alberto Katsumiti
title	Immortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro
title_short	Immortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro
title_full	Immortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro
title_fullStr	Immortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro
title_full_unstemmed	Immortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro
title_sort	immortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/937c8d854bab44f99579b0458b77380f
work_keys_str_mv	AT albertokatsumiti immortalisationofprimaryhumanalveolarepitheliallungcellsusinganonviralvectortostudyrespiratorybioreactivityinvitro AT pakatipruenraroengsak immortalisationofprimaryhumanalveolarepitheliallungcellsusinganonviralvectortostudyrespiratorybioreactivityinvitro AT mirenpcajaraville immortalisationofprimaryhumanalveolarepitheliallungcellsusinganonviralvectortostudyrespiratorybioreactivityinvitro AT andrewjthorley immortalisationofprimaryhumanalveolarepitheliallungcellsusinganonviralvectortostudyrespiratorybioreactivityinvitro AT teresadtetley immortalisationofprimaryhumanalveolarepitheliallungcellsusinganonviralvectortostudyrespiratorybioreactivityinvitro
_version_	1718395595137220608

Immortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro

Ejemplares similares