Improvement on primary culture of human nasal epithelium by enzymatical dissociation.
- Author:
Jihong YANG
1
;
Gehua ZHANG
;
Yan WEI
;
Yuan LI
Author Information
1. Department of Otolaryngology-Head and Neck Surgery, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, 510630, China.
- Publication Type:Journal Article
- MeSH:
Cell Culture Techniques;
methods;
Cell Division;
Cells, Cultured;
Epithelial Cells;
cytology;
Humans;
Nasal Mucosa;
cytology
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2009;23(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 confluence 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.
