Improvement on primary culture of human nasal epithelium by enzymatical dissociation
10.3969/j.issn.1001-1781.2009.23.004
- VernacularTitle:酶消化分离法原代培养人鼻黏膜上皮细胞的改进
- Author:
Jihong YANG
;
Gehua ZHANG
;
Yan WEI
;
Yuan LI
- Publication Type:Journal Article
- Keywords:
nasal mucosa;
epithelium;
cell culture
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2009;(23):1066-1068
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To highlight the key points of primary culture of human nasal epithelial cells by enzymatical dissociation for high achievement ratio,and to establish a successful primary culture model for subsequent experiments.Method:Primary culture of human nasal epithelial cells was performed with enzymatical dissociation of isolated tissue in serum-free medium. On the basis of this method,some improvements were subjected,such as stripping mucosal epithelium from adjacent connective tissue,applying DNase type Ⅰ to digesting procedure,adding trypsin directly to the collagenase solution containing digested mucosa pieces,employing uncoated culture dishes and so on. Immunofluorescence with a monoclonal anti-cytokeratin antibody 8/18 was used to confirm the epithelial nature of the cultured cells. Result:Nasal epithelial cells grew well and confluenced on the 6th to 8th day. Positive expression of cytokeratin(CK)8/18 showed the epithelial property of cultured cells.Conclusion:Primary culture model of human nasal epithelial cells can be successfully established by enzymatical dissociation. Improvements on processes of material using and enzyme digestion can gain a high achievement ratio and harvest a high purity and certain amount of reliable primary epithelial cells.
