Successful establishment of primary small airway cell cultures in human lung transplantation.
BACKGROUND
The study of small airway diseases such as post-transplant bronchiolitis obliterans syndrome (BOS) is hampered by the difficulty in assessing peripheral airway function either physiologically or directly. Our aims were to develop robust methods for sampling small airway epithelial cells (SAEC) and to establish submerged SAEC cultures for downstream experimentation.


METHODS
SAEC were obtained at 62 post-transplant bronchoscopies in 26 patients using radiologically guided bronchial brushings. Submerged cell cultures were established and SAEC lineage was confirmed using expression of clara cell secretory protein (CCSP).


RESULTS
The cell yield for SAEC (0.956 +/- 0.063 x 106) was lower than for large airway cells (1.306 +/- 0.077 x 106) but did not significantly impact on the culture establishment rate (79.0 +/- 5.2% vs. 83.8 +/- 4.7% p = 0.49). The presence of BOS significantly compromised culture success (independent of cell yield) for SAEC (odds ratio (95%CI) 0.067 (0.01-0.40)) but not LAEC (0.3 (0.05-1.9)). Established cultures were successfully passaged and expanded.


CONCLUSIONS
Primary SAEC can be successfully obtained from human lung transplant recipients and maintained in culture for downstream experimentation. This technique will facilitate the development of primary in vitro models for BOS and other diseases with a small airway component such as asthma, cystic fibrosis and COPD.
Queensland Centre for Pulmonary Transplantation and Vascular Disease, The Prince Charles Hospital, Brisbane, 4032, Queensland, Australia. bbanerjee@ichr.uwa.edu.au
Balarka
B
Banerjee
Anthony
A
Kicic
Michael
M
Musk
Erika N
EN
Sutanto
Stephen M
SM
Stick
Daniel C
DC
Chambers
D000293:Adolescent; D000328:Adult; D001989:Bronchiolitis Obliterans / Q000209:etiology / Q000473:pathology*; D001999:Bronchoscopy; D018929:Cell Culture Techniques; D019070:Cell Lineage; D049109:Cell Proliferation; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism / Q000473:pathology*; D005260:Female; D006801:Humans; D008168:Lung / Q000473:pathology / Q000601:surgery*; D016040:Lung Transplantation / Q000009:adverse effects; D008297:Male; D008875:Middle Aged; D016017:Odds Ratio; D014598:Uteroglobin / Q000378:metabolism
C556389:SCGB1A1 protein, human; D014598:Uteroglobin