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Updated: Sep 27, 2026
Isolation and Culture of Primary Human Gingival Epithelial Cells using Y-27632
Published on: November 6, 2021
[Research on inhibition of areca nut extract on human buccal keratinocytes]
Objective:
To culture human oral mucosa epithelial cells in vitro and study the mechanism of changes in the epithelia of oral submucous fibrosis (OSF).
Methods:
The keratinocytes were obtained from normal buccal mucosa (NM-KC) and OSF buccal mucosa (OSF-KC) with dispase trypsine and cultured in vitro. Then the cell proliferation of keratinocytes incubated with or without areca nut extract in the presence of 10% fetal calf serum for 48 hours at 37 degrees C in 5% CO2 and air were monitored by use of thiazolyl blue (MTT) assay.
Results:
Primary culture grew in small islands that coalesced at confluency. Ultrastructurally the cells contained a large of tonofilament and microvilli. Immunohistochemistry demonstrated uniform staining of the cells with antibodies to keratins. There was no difference in cell proliferation between OSF-KC and NM-KC, and areca nut extract inhibited keratinocyte proliferation in a concentration-dependent manner.
Conclusion:
The results suggested that utilizing dispase II can establish fibroblast-free culture for growing of human oral mucosa keratinocytes and areca nut extract is a cytotoxic agent to human buccal keratinocytes.
