Non-bronchoscopic sampling and culture of bronchial epithelial cells in children.
BACKGROUND
The bronchial epithelium is likely to play a vital role in airway diseases in children, such as asthma and viral-associated wheeze. In adults, studies with primary bronchial epithelial cells cultured from samples obtained by fibre-optic bronchoscopy have provided key insights into the role of the epithelial cell. However, it is difficult to justify bronchoscopy in children to obtain epithelial cells for research purposes.


OBJECTIVE
To examine the possibility of retrieving and culturing viable epithelial cells using a blind non-bronchoscopic method from children undergoing elective surgery.


METHODS
Subjects were children undergoing elective surgery under general anaesthesia. Following intubation, non-bronchoscopic bronchoalveolar lavage and non-bronchoscopic bronchial brushing were performed. A sheathed bronchial cytology brush was advanced through the endotracheal tube, wedged and then withdrawn 2-3 cm before gentle sampling was used to collect bronchial epithelial cells. Initial samples were used to characterize the number, type and viability of epithelial cells recovered compared to a control group of adults undergoing standard bronchoscopic sampling. Subsequent samples were used to establish primary bronchial epithelial cell cultures in children both with and without wheezing illness.


RESULTS
A total of 63 children underwent bronchial brushing [38 male; median age 7.1 years (1.0-14.2 years]. Initial samples (n=30) showed recovery of viable epithelial cells comparable to that from a single brush obtained via a bronchoscope in an adult control group (n=11). In 27 (82%) of the subsequent 33 samples obtained non-bronchoscopically from children, primary bronchial epithelial cell cultures were successfully established. There were no adverse effects attributable to sampling.


CONCLUSIONS
We have shown that non-bronchoscopic bronchial brushing is a safe and effective technique for recovering viable bronchial epithelial cells that consistently yield primary cultures. This method will facilitate examination of the role of the epithelium in paediatric disease.
Doherty
G M
GM
Department of Child Health, Queen's University, Belfast, Northern Ireland.
Christie
S N
SN
Skibinski
G
G
Puddicombe
S M
SM
Warke
T J
TJ
de Courcey
F
F
Cross
A L
AL
Lyons
J D M
JD
Ennis
M
M
Shields
M D
MD
Heaney
L G
LG
D000293:Adolescent; D001249:Asthma / Q000175:diagnosis / Q000473:pathology; D001980:Bronchi / Q000473:pathology*; D018893:Bronchoalveolar Lavage / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003581:Cytodiagnosis / Q000379:methods; D004847:Epithelial Cells / Q000473:pathology*; D005240:Feasibility Studies; D005260:Female; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007223:Infant; D007633:Keratins / Q000032:analysis; D008297:Male; D012135:Respiratory Sounds / Q000175:diagnosis; D013048:Specimen Handling / Q000379:methods
D007633:Keratins