Toggle Main Menu Toggle Search

Open Access padlockePrints

Isolation and characterisation of human pulmonary microvascular endothelial cells from patients with severe emphysema

Lookup NU author(s): Dr Laura Mackay, Dr James Lordan, Professor Andrew FisherORCiD, Emeritus Professor Nick Europe-Finner


Full text for this publication is not currently held within this repository. Alternative links are provided below where available.


Background: Loss of the pulmonary microvasculature in the pathogenesis of emphysema has been put forward as a credible alternative to the classical inflammatory cell driven proteolysis hypothesis. Mechanistic studies in this area have to date employed animal models, immortalised cell lines, primary endothelial cells isolated from large pulmonary arteries and non-pulmonary tissues and normal human pulmonary microvascular endothelial cells. Although these studies have increased our understanding of endothelial cell function, their relevance to mechanisms in emphysema is questionable. Here we report a successful technique to isolate and characterise primary cultures of pulmonary microvascular endothelial cells from individuals with severe emphysema. Methods: A lobe of emphysematous lung tissue removed at the time of lung transplantation surgery was obtained from 14 patients with severe end-stage disease. The pleura, large airways and large blood vessels were excised and contaminating macrophages and neutrophils flushed from the peripheral lung tissue before digestion with collagenase. Endothelial cells were purified from the cell mixture via selection with CD31 and UEA-1 magnetic beads and characterised by confocal microscopy and flow cytometry. Results: Successful isolation was achieved from 10 (71%) of 14 emphysematous lungs. Endothelial cells exhibited a classical cobblestone morphology with high expression of endothelial cell markers (CD31) and low expression of mesenchymal markers (CD90, alpha SMA and fibronectin). E-selectin (CD62E) was inducible in a proportion of the endothelial cells following stimulation with TNF alpha, confirming that these cells were of microvascular origin. Conclusions: Emphysematous lungs removed at the time of transplantation can yield large numbers of pulmonary microvasculature endothelial cells of high purity. These cells provide a valuable research tool to investigate cellular mechanisms in the pulmonary microvasculature relevant to the pathogenesis of emphysema.

Publication metadata

Author(s): Mackay LS, Dodd S, Dougall IG, Tomlinson W, Lordan J, Fisher AJ, Corris PA

Publication type: Article

Publication status: Published

Journal: Respiratory Research

Year: 2013

Volume: 14

Pages: 23

Print publication date: 01/02/2013

ISSN (print): 1465-9921

ISSN (electronic): 1465-993X

Publisher: BioMed Central Ltd.


DOI: 10.1186/1465-9921-14-23

Notes: Article no. 23 is 11 pp.


Altmetrics provided by Altmetric