Influencing Factors of Short-term Exposure Test for Rabbit Corneal Epithelial Cells

Objectives  To observe the influencing factors of short-term exposure test (STE test) in rabbit corneal epithelial cells.

Methods  Statens Seruminstitut Rabbit Cornea cells were seeded at the initial density of 3×10³ and 6×10³ cells/well respectively. 12 chemicals, including benzalkonium chloride, were exposed to cultured cells at the doses of 5% and 0.05% respectively for 5 minutes, and cell viability were detected by MTT assay and cck-8 assay, The eye irritation of the above chemicals was evaluated by the traditional method and the STE integration method.

Results  Seeded at 3×10³ cells/well and 6×10³ cells/well, the cell viability result were correlated well after exposure. The cell viability result measured by MTT assay and CCK-8 assay were significantly correlated. The traditional method and STE integration method were consistented with the result of grading judgments for severely irritating substances and mild irritants.

Conclusions  The initial cell inoculation density, different color method and result criteria were the influencing factors of short-term exposure test of rabbit corneal epithelial cells. The density of 3×10³ or 6×10³ cells/well, the CCK-8 assay for cell viability determination, and the STE integration method for the eye irritation evaluation could be optional for the experiment.