Chest
Volume 138, Issue 6, December 2010, Pages 1441-1447
Journal home page for Chest

Original Research
Inherited Lung Diseases
Ciliated Air-Liquid Cultures as an Aid to Diagnostic Testing of Primary Ciliary Dyskinesia

https://doi.org/10.1378/chest.10-0175Get rights and content

Background

The diagnosis of primary ciliary dyskinesia (PCD) can prove difficult because of secondary damage of ciliated tissue.

Methods

Here we audit culturing cells, obtained by nasal brushing, to a ciliated phenotype using an air-liquid interface method to determine if the effects of secondary damage on cilia were reduced following culture.

Results

Of 231 patients consecutively referred for diagnostic testing, culture was attempted in 187, with 101 (54%) becoming ciliated. Of the 90 brush biopsy samples with a low dyskinesia score (< 40%), 71 grew cilia after culture (79% success). Significant secondary damage (> 40% dyskinesia) was present in 69 (43%) of the initial brush biopsy samples, and of these, 18 (26%) became ciliated after culture. In these samples, ciliary dyskinesia was significantly (P < .001) reduced (64% ± 6.8% before culture, 31% ± 4.5% after culture). Ciliary beat frequency (CBF) after cell culture was similar to CBF before culture. Cell culture helped to exclude PCD in eight patients for whom ciliary dyskinesia was present in > 70% of the initial brush biopsy sample, a level at which a rebiopsy would normally be requested. In six patients in whom no cilia were found in the initial brush biopsy samples, ciliated cell culture was successful and excluded the diagnosis. PCD was diagnosed in 28 patients and ciliated cell culture was successful in 12 (43%) showing identical ciliary beat pattern and electron microscopy findings.

Conclusions

Ciliary dyskinesia was reduced following cell culture to a ciliated phenotype compared with the initial brush biopsy sample. The specific PCD phenotype was maintained after culture.

Section snippets

Patients

Between April 1, 2007, and March 31, 2008, 231 patients were referred to the PCD diagnostic service in Leicester. This center is one of the three national PCD diagnostic centers in England, all of which are funded to carry out cell culture routinely on clinical samples. This report is an audit of results of air-liquid interface cultures of patients referred for diagnostic testing of PCD in Leicester. All patients gave their consent for participating in the PCD service, and all samples were

Results

Between April 1, 2007, and March 31, 2008, 231 patients were referred for diagnostic testing for PCD. Of the patients referred, 217 had a brush biopsy performed.

Discussion

Although we found cell culture with redifferentiation of the ciliated epithelium helpful in a number of patients, it is a highly specialized and time-consuming method. A major advantage was that, when successful, this technique allowed detailed evaluation of both ciliary beat pattern and electron microscopy. In our hands only 54% of biopsy samples could be cultured to a ciliated phenotype. This is likely to be due to a number of factors. First, our brushing technique obtains relatively few

Acknowledgments

Author contributions: Dr Hirst explicitly declares that he has had access to and takes responsibility for the integrity of the data and the accuracy of the data analysis.

Dr Hirst: contributed to writing the manuscript, analyzing the data, and developing the cell culture method.

Mr Rutman: contributed to performing the electron microscopy.

Ms Williams: contributed to culturing the cells.

Dr O'Callaghan: contributed to preparing the manuscript and is the clinical leader of the PCD service in

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    Funding/Support: This study was funded by the National Commissioning Group via the National Health Service, England.

    Reproduction of this article is prohibited without written permission from the American College of Chest Physicians (http://www.chestpubs.org/site/misc/reprints.xhtml).

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