Human nasal turbinates as a viable source of respiratory epithelial cells using co-culture system versus dispase-dissociation technique.
OBJECTIVE
To compare a co-culture system with a conventional dispase-dissociation method for obtaining functional human respiratory epithelial cells from the nasal turbinates for tissue engineering application.


METHODS
Human respiratory epithelial cells were serially passaged using a co-culture system and a conventional dispase-dissociation technique. The growth kinetics and gene expression levels of the cultured respiratory epithelial cells were compared. Four genes were investigated, namely cytokeratin-18, a marker for ciliated and secretory epithelial cells; cytokeratin-14, a marker for basal epithelial cells; MKI67, a proliferation marker; and MUC5B, a marker for mucin secretion. Immunocytochemical analysis was performed using monoclonal antibodies against the high molecular-weight cytokeratin 34 beta E12, cytokeratin 18, and MUC5A to investigate the protein expression from cultured respiratory epithelial cells.


RESULTS
Respiratory epithelial cells cultured using both methods maintained polygonal morphology throughout the passages. At passage 1, co-cultured respiratory epithelial showed a 2.6-times higher growth rate compared to conventional dispase dissociation technique, and 7.8 times higher at passage 2. Better basal gene expression was observed by co-cultured respiratory epithelial cells compared to dispase dissociated cells. Immunocytochemical analyses were positive for the respiratory epithelial cells cultured using both techniques.


CONCLUSIONS
Co-culture system produced superior quality of cultured human respiratory epithelial cells from the nasal turbinates as compared to dispase dissociation technique.
Noruddin
Nur Adelina Ahmad
NA
Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia (National University of Malaysia), Ampang Puteri Specialist Hospital, Malaysia.
Saim
Aminuddin B
AB
Chua
Kien Hui
KH
Idrus
Ruszymah
R
D015415:Biomarkers / Q000378:metabolism; D048708:Cell Growth Processes / Q000235:genetics; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D004247:DNA / Q000235:genetics; D010450:Endopeptidases; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D007158:Immunologic Techniques; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D053547:Keratin-14 / Q000096:biosynthesis / Q000235:genetics; D053538:Keratin-18 / Q000096:biosynthesis / Q000235:genetics; D055271:Mucin 5AC; D009077:Mucins / Q000096:biosynthesis / Q000235:genetics; D009297:Nasal Mucosa / Q000166:cytology*; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D023822:Tissue Engineering / Q000379:methods; D014420:Turbinates / Q000166:cytology*
D015415:Biomarkers; D047908:Intracellular Signaling Peptides and Proteins; D053547:Keratin-14; D053538:Keratin-18; C524678:MUC5AC protein, human; D055271:Mucin 5AC; D009077:Mucins; C492406:NIFK protein, human; D009687:Nuclear Proteins; D004247:DNA; D010450:Endopeptidases; C023139:dispase