MTT比色法检测兔角膜上皮细胞活性

MTT colorimetric assay of corneal epithelial cell activity

目的 :建立一种定量检测角膜上皮细胞活性的方法。方法 :造成兔角膜碱烧伤和激光烧伤模型 ,分别于伤后 1,2 ,3周钻取板层角膜 ,37℃孵育 1h ,加MTT孵育 4h ,加DMSO充分溶解 ,于酶联免疫仪上 4 90nm处测定D值。结果 :经MTT比色法测定 ,碱烧伤和激光烧伤后不同时间点的兔角膜上皮细胞活性均明显低于正常对照组 (P <0 .0 1)。结论 :MTT比色法可用于定量检测角膜上皮细胞活性 。

Objective:To establish a method for measuring the activity of cornea epithelium quantitatively. Methods:Rabbit corneas were burnt either by alkali or by CO 2 laser. The lamellar cornea was cut at the end of 1,2 and 3 weeks and cultured in 2 ml DMEM with 5% CO 2, 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. [