机构:[1]Department of Otolaryngology, Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100005, China临床科室耳鼻咽喉-头颈外科首都医科大学附属北京同仁医院首都医科大学附属同仁医院[2]Key Laboratory of Otolaryngology, Head and Neck Surgery (Ministry of Education), Beijing Institute of Otolaryngology, Beijing 100005, China研究所耳鼻咽喉科研究所首都医科大学附属北京同仁医院首都医科大学附属同仁医院
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 beta-tubulin IVand ZO-1 immunocytochemistry. Basal and ATP-stimulated ciliary beat frequency (CBF) was measured using a highspeed 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 mu M 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.
基金:
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).
第一作者机构:[1]Department of Otolaryngology, Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100005, China[2]Key Laboratory of Otolaryngology, Head and Neck Surgery (Ministry of Education), Beijing Institute of Otolaryngology, Beijing 100005, China
通讯作者:
通讯机构:[1]Department of Otolaryngology, Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing 100005, China[2]Key Laboratory of Otolaryngology, Head and Neck Surgery (Ministry of Education), Beijing Institute of Otolaryngology, Beijing 100005, China
推荐引用方式(GB/T 7714):
Jino Jian,Meng Na,Wang Hong,et al.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.doi:10.1007/s11684-013-0287-x.
APA:
Jino, Jian,Meng, Na,Wang, Hong&Zhang, Luo.(2013).Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro.FRONTIERS OF MEDICINE,7,(4)
MLA:
Jino, Jian,et al."Comparison of human nasal epithelial cells grown as explant outgrowth cultures or dissociated tissue cultures in vitro".FRONTIERS OF MEDICINE 7..4(2013):486-491