[Improvement on primary culture of human nasal epithelium by enzymatical dissociation].

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.