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噻唑蓝比色法测定待移植睾丸细胞的活性
Determination of pretransplantational cell viability by MTT colorimetric assay.
【摘要】 目的 探讨噻唑蓝( MTT) 比色法测定细胞线粒体琥珀酸脱氢酶, 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MTTformazan , 上清液于563 nm 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 基质浓度1 g/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 法较台盼蓝法灵敏、准确。应用MTT 技术为10 例人睾丸Leydig 细胞移植术监测细胞活性获得成功。结论 MTT 比色法为细胞移植术提供了一种监测细胞活性的客观、准确方法。
【Abstract】 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.
【Key words】 MTT colorimetric assay Cell/viability Cell/transplantation Trypan bluedye;
- 【文献出处】 中华器官移植杂志 ,CHINEAE JOURNAL OF ORGAN TRANSPLANTATION , 编辑部邮箱 ,1999年04期
- 【分类号】R619
- 【被引频次】1
- 【下载频次】61