MTT colorimetric assay of corneal epithelial cell activity / 军事医学科学院院刊

ABSTRACT

Objective:To establish a method for measuring the activity of cornea epithelium quantitatively. Methods:Rabbit corneas were burnt either by alkali or by CO2 laser. The lamellar cornea was cut at the end of 1,2 and 3 weeks and cultured in 2 ml DMEM with 5% CO2, 37℃ for 1 h.Then 200 μl of MTT was added to the culture followed by incubation for another 4 h. The supernatant was discarded and 4 ml of DMSO was added into each culturedish for dissolving MTT completely under the condition of room temperature.200 μl of DMSO sample was added to each well of 96-well plate and each sample was triplicated. The absorbance of the plate was measured at 490 nm ultraviolet. Results:The D value of the burnt corneas was obviously lower than that of the normal ones(P<0.01). Conclusion:MTT method can be used to measure the activity of cornea epithelium quantitatively.