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

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PDF(183 KB)
Front. Med. ›› 2013, Vol. 7 ›› Issue (4) : 486-491. DOI: 10.1007/s11684-013-0287-x
RESEARCH ARTICLE

Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro

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Abstract

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.

Keywords

ciliated cells / ciliary beat frequency / dissociated tissue culture / explant culture / nasal epithelial cells

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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 https://doi.org/10.1007/s11684-013-0287-x

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Acknowledgements

The authors thank Dr. Jagdish Devalia (JD Medical and Scientific Communications, Southall, UK) for help with editing this manuscript. This work was supported by the National Science Fund for Distinguished Young Scholars (Grant No. 81025007), National Natural Science Foundation of China (Grant Nos. 81100704 and 30973282), Beijing Natural Science Foundation (7131006), Ministry of Health Foundation (201202005), Beijing Nova Program (Z111107054511061), Specialized Research Fund for the Doctoral Program of Higher Education of China (20111107120004), The Capital Health Research and Development of Special (2011-1017-03), and Science Foundation for High-Level Medical Talents of Beijing Health System (2009-02-007).
Compliance with ethics guidelines
Jian Jiao, Na Meng, Hong Wang, and Luo Zhang declare that they have no conflict of interest. All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000 (5). Informed consent was obtained from all patients for providing the nasal tissues samples.

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2014 Higher Education Press and Springer-Verlag Berlin Heidelberg
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