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Human Nasal Turbinates as a Viable Source of Respiratory Epithelial Cells Using Co‐Culture System Versus Dispase Dissociation Technique
Author(s) -
Noruddin Nur Adelina Ahmad,
Saim Aminuddin B.,
Chua Kien Hui,
Idrus Ruszymah
Publication year - 2007
Publication title -
the laryngoscope
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.181
H-Index - 148
eISSN - 1531-4995
pISSN - 0023-852X
DOI - 10.1097/mlg.0b013e3181453a1e
Subject(s) - dispase , cytokeratin , respiratory system , pathology , epithelium , biology , cell culture , respiratory epithelium , microbiology and biotechnology , medicine , immunohistochemistry , anatomy , biochemistry , enzyme , genetics , collagenase
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 34βE12, 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. Conclusion: Co‐culture system produced superior quality of cultured human respiratory epithelial cells from the nasal turbinates as compared to dispase dissociation technique.