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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。结论ＭＴＴ比色法可用于分析ＲＰＥ增生活性和测定抗代谢药物的细胞毒作用
Objective To investigate the feasibility of using MTT colorimetric assay in the study of retinal pigment epithelium (RPE) cells. Methods The experimental conditions of MTT were screened one by one. The cytotoxic effect of daunorubicin on RPE was compared with the cell counting method and the clonogenic assay. Results MTT effect more than 6h, dimethyl sulfoxide dissolved 5 ~ 10min immediately after the determination of absorbance (A value) is the most appropriate conditions. A550nm is greater than A490nm, there is no significant difference between the two wavelengths (P> 0.05). The number of cells was linearly related to A (P <0.01). The EC50 of daunorubicin on RPE cells by MTT assay, cell counting method and clonogenic assay were 20.00, 1615 and 1224 μg / L, respectively. The EC90 values were 118.02, 145.59 and 42, respectively. 50 μg / L. Conclusion MTT assay can be used to analyze the proliferative activity of RPE and determine the cytotoxicity of antimetabolites