Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro
Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro
Jian Jiao, Na Meng, Hong Wang, Luo Zhang()
Department of Otolaryngology, Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100005, China; Key Laboratory of Otolaryngology, Head and Neck Surgery (Ministry of Education), Beijing Institute of Otolaryngology, Beijing 100005, China
The purpose of this study was to compare cell growth characteristics, ciliated cell differentiation, and function of human nasal epithelial cells established as explant outgrowth cultures or dissociated tissue cultures. Human nasal mucosa of the uncinate process was obtained by endoscopy and epithelial cell cultures were established by explant outgrowth or dissociated tissue culture methods. Epithelial cell growth characteristics were observed by inverted phase contrast microscopy. Ciliated cell differentiation was detected by β-tubulin IV and ZO-1 immunocytochemistry. Basal and ATP-stimulated ciliary beat frequency (CBF) was measured using a high-speed digital microscopic imaging system. Both the explant and dissociated tissue cultures established as monolayers with tight junctions and differentiated cell composition, with both types of cultures comprising ciliated and non-ciliated epithelial cells. Fibroblasts were also frequently found in explant cultures but rarely seen in dissociated tissue cultures. In both culture systems, the highest ciliated cell density appeared at 7th–10th culture day and declined with time, with the lifespan of ciliated cells ranging from 14 to 21 days. Overall, 10% of the cells in explant cultures and 20% of the cells in the dissociated tissue cultures were ciliated. These two cultures demonstrated similar ciliary beat frequency values at baseline (7.78±1.99 Hz and 7.91±2.52 Hz, respectively) and reacted equivalently following stimulation with 100 μM ATP. The results of this study indicate that both the explant outgrowth and dissociated tissue culture techniques are suitable for growing well-differentiated nasal ciliated and non-ciliated cells, which have growth characteristics and ciliary activity similar to those of nasal epithelial cells in vivo.
. Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro[J]. Frontiers of Medicine, 2013, 7(4): 486-491.
Jian Jiao, Na Meng, Hong Wang, Luo Zhang. Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro. Front Med, 2013, 7(4): 486-491.
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