噻唑蓝比色法测定待移植睾丸细胞的活性

目的 探讨噻唑蓝（ＭＴＴ）比色法测定细胞线粒琥珀酸脱氢酶，建立细胞活性的比色测定法。方法 采用大鼠睾丸制备的Ｌｅｙｄｉｇ细胞悬液，与ＭＴＴ基质液共同温育后，以异丙醇溶解ＭＴＴ ｆｏｒｍａｚａｎ，上清液于５６３ｎｍ测吸光度；用体外细胞培养和睾酮释放量评价方法的可靠性，与台盼蓝染色法对照。结果 基质深度１ｇ／Ｌ，温育时间３小时，异丙醇作溶剂为反应最佳条件。细胞培养和睾酮测定表明ＭＴＴ法中盼蓝法灵敏、准

Determination of pretransplantational cell viability by MTT colorimetric assay

Objective To assess the role of MTT colorimetric assay in measuring mitochondrial succinate dehydrogenase (SDH) and establish a new way to determine pretransplantational cell viability.Methods Human or rat testicular Leydig cell suspensions were incubated with MTT medium, isopropanol was used to dissolve MTT formazan, then the absorbance of supertants was measured at 563?nm wavelength. Cell culture and testosterone production were used to assess the reliability of MTT colorimetric assay with comparison with Trypan bluedye.Results 1 g/L MTT concentration, 3?h incubation, and isopropanol as a solvent were the best optimum for the cell culture in vitro and testosterone determination showed that MTT colorimetric assay was more sensitive and accurate than Trypan bluedye. MTT colorimetric assay was employed to successfully measure cell viability for 10 cases of human testicular Leydig cell transplantation.Conclusion MTT colorimetric assay provided a reliable, objective, accurate method for determining the pretransplantational cell viability.