摘要
目的探讨ＭＴＴ比色法用于视网膜色素上皮（ＲＰＥ）细胞研究的可行性。方法逐项筛选ＭＴＴ法的实验条件，与细胞计数法和克隆形成试验比较，观察柔红霉素对ＲＰＥ的细胞毒作用。结果ＭＴＴ作用６ｈ以上、二甲基亚砜溶解５～１０ｍｉｎ后即刻测定吸光度（Ａ值）是最适宜的条件。Ａ５５０ｎｍ大于Ａ４９０ｎｍ，由两种波长所得抑制率间无显著差异（Ｐ＞０．０５）。细胞数与Ａ值成直线相关（Ｐ＜０．０１）。由ＭＴＴ法、细胞计数法和克隆形成试验所得柔红霉素对ＲＰＥ细胞的ＥＣ５０分别为２０．００、１６１５和１２２４μｇ／Ｌ，ＥＣ９０分别为１１８．０２、１４５．５９和４２．５０μｇ／Ｌ。
ObjectiveTo investigate the feasibility in a tetrazolium colorimetric assay(MTT assay)applied for retinal pigment epithelial(RPE)cells.MethodsThe optimal variables in MTT assay were selected one by one.Compared with cell counting and clone formation test,MTT assay was used in study of the cytotoxicity of daunoblastina on RPE cells.ResultsThe optimal variables for RPE cells were more than 6 h for MTT incubation time,formazan dissolved in DMSO for 5～10 min and immediate measurement of the solution.Absorbance(A value)at 550 nm was greater than that at 490 nm,but there was no statistical difference between the two kinds of filters when compared by percentage of cellular inhibition.A linear relationship was noted between A value and RPE cell number.The concentration of daunoblastina required for 50% growth inhibition of RPE cells by MTT assay,cell counting and clone formation test were 20.00,16.15 and 12.24 μg/L.The concentration required for 90% growth inhibition of RPE cells were 118.02 , 145.59 and 42.50 μg/L,respectively.ConclusionMTT assay is valuable for measuring RPE cell proliferation and cytotoxicity of antimetablic agents.
出处
《眼科研究》
CSCD
1998年第2期102-104,共3页
Chinese Ophthalmic Research
基金
国家自然科学基金
