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Long term culture of the A549 cancer cell line promotes multilamellar body formation and differentiation towards an Alveolar type II pneumocyte phenotype

Long term culture of the A549 cancer cell line promotes multilamellar body formation and differentiation towards an Alveolar type II pneumocyte phenotype
Long term culture of the A549 cancer cell line promotes multilamellar body formation and differentiation towards an Alveolar type II pneumocyte phenotype

Pulmonary research requires models that represent the physiology of alveolar epithelium but concerns with reproducibility, consistency and the technical and ethical challenges of using primary or stem cells has resulted in widespread use of continuous cancer or other immortalized cell lines. The A549 'alveolar' cell line has been available for over four decades but there is an inconsistent view as to its suitability as an appropriate model for primary alveolar type II (ATII) cells. Since most work with A549 cells involves short term culture of proliferating cells, we postulated that culture conditions that reduced proliferation of the cancer cells would promote a more differentiated ATII cell phenotype. We examined A549 cell growth in different media over long term culture and then used microarray analysis to investigate temporal regulation of pathways involved in cell cycle and ATII differentiation; we also made comparisons with gene expression in freshly isolated human ATII cells. Analyses indicated that long term culture in Ham's F12 resulted in substantial modulation of cell cycle genes to result in a quiescent population of cells with significant up-regulation of autophagic, differentiation and lipidogenic pathways. There were also increased numbers of up- and down-regulated genes shared with primary cells suggesting adoption of ATII characteristics and multilamellar body (MLB) development. Subsequent Oil Red-O staining and Transmission Electron Microscopy confirmed MLB expression in the differentiated A549 cells. This work defines a set of conditions for promoting ATII differentiation characteristics in A549 cells that may be advantageous for studies with this cell line.

A549 Cells, Alveolar Epithelial Cells, Cell Culture Techniques, Cell Cycle, Cell Differentiation, Gene Expression Regulation, Humans, Microscopy, Electron, Transmission, Oligonucleotide Array Sequence Analysis, Phenotype, Polymerase Chain Reaction, Journal Article
1932-6203
e0164438
Cooper, James Ross
17051c51-b879-400f-aae3-0f326868a582
Abdullatif, Muhammad Bilal
27219314-13a2-484c-9f85-14404d3275f0
Burnett, Edward C
8f36a8fb-c919-4f04-9b53-afa1cb57be28
Kempsell, Karen E
46bb6be3-c900-498e-8728-66b2ffcc9e38
Conforti, Franco
28bf123c-e42a-4fb5-8b26-f79e1095c586
Tolley, Howard
7633f424-646e-4dfe-90a1-90884bb9b453
Collins, Jane E
be0e66f1-3036-47fa-9d7e-914c48710ba4
Davies, Donna E
7de8fdc7-3640-4e3a-aa91-d0e03f990c38
Cooper, James Ross
17051c51-b879-400f-aae3-0f326868a582
Abdullatif, Muhammad Bilal
27219314-13a2-484c-9f85-14404d3275f0
Burnett, Edward C
8f36a8fb-c919-4f04-9b53-afa1cb57be28
Kempsell, Karen E
46bb6be3-c900-498e-8728-66b2ffcc9e38
Conforti, Franco
28bf123c-e42a-4fb5-8b26-f79e1095c586
Tolley, Howard
7633f424-646e-4dfe-90a1-90884bb9b453
Collins, Jane E
be0e66f1-3036-47fa-9d7e-914c48710ba4
Davies, Donna E
7de8fdc7-3640-4e3a-aa91-d0e03f990c38

Cooper, James Ross, Abdullatif, Muhammad Bilal, Burnett, Edward C, Kempsell, Karen E, Conforti, Franco, Tolley, Howard, Collins, Jane E and Davies, Donna E (2016) Long term culture of the A549 cancer cell line promotes multilamellar body formation and differentiation towards an Alveolar type II pneumocyte phenotype. PLoS ONE, 11 (10), e0164438. (doi:10.1371/journal.pone.0164438).

Record type: Article

Abstract

Pulmonary research requires models that represent the physiology of alveolar epithelium but concerns with reproducibility, consistency and the technical and ethical challenges of using primary or stem cells has resulted in widespread use of continuous cancer or other immortalized cell lines. The A549 'alveolar' cell line has been available for over four decades but there is an inconsistent view as to its suitability as an appropriate model for primary alveolar type II (ATII) cells. Since most work with A549 cells involves short term culture of proliferating cells, we postulated that culture conditions that reduced proliferation of the cancer cells would promote a more differentiated ATII cell phenotype. We examined A549 cell growth in different media over long term culture and then used microarray analysis to investigate temporal regulation of pathways involved in cell cycle and ATII differentiation; we also made comparisons with gene expression in freshly isolated human ATII cells. Analyses indicated that long term culture in Ham's F12 resulted in substantial modulation of cell cycle genes to result in a quiescent population of cells with significant up-regulation of autophagic, differentiation and lipidogenic pathways. There were also increased numbers of up- and down-regulated genes shared with primary cells suggesting adoption of ATII characteristics and multilamellar body (MLB) development. Subsequent Oil Red-O staining and Transmission Electron Microscopy confirmed MLB expression in the differentiated A549 cells. This work defines a set of conditions for promoting ATII differentiation characteristics in A549 cells that may be advantageous for studies with this cell line.

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More information

Accepted/In Press date: 26 September 2016
e-pub ahead of print date: 28 October 2016
Published date: 28 October 2016
Keywords: A549 Cells, Alveolar Epithelial Cells, Cell Culture Techniques, Cell Cycle, Cell Differentiation, Gene Expression Regulation, Humans, Microscopy, Electron, Transmission, Oligonucleotide Array Sequence Analysis, Phenotype, Polymerase Chain Reaction, Journal Article

Identifiers

Local EPrints ID: 414178
URI: https://eprints.soton.ac.uk/id/eprint/414178
ISSN: 1932-6203
PURE UUID: b0c18cb5-cc03-4ba3-a621-f5a2372e85f2
ORCID for Donna E Davies: ORCID iD orcid.org/0000-0002-5117-2991

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Date deposited: 15 Sep 2017 16:31
Last modified: 03 Dec 2019 02:07

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Contributors

Author: James Ross Cooper
Author: Muhammad Bilal Abdullatif
Author: Edward C Burnett
Author: Karen E Kempsell
Author: Franco Conforti
Author: Howard Tolley
Author: Jane E Collins
Author: Donna E Davies ORCID iD

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