U-SENS™试验方法的建立与探讨

Establishment and Discussion of U-SENSTM Test Method

关键词:  U937  流式细胞术  化妆品替代

Key words:  U937  flow cytometry  the substitute methods for cosmetics

摘要:

目的 建立符合中国实际情况的U-SENS^™试验方法，并验证新试验方法的可靠性。方法 参考OECD指导原则442E附录II U-SENS^™试验方法，根据其中的QC模板，建立以不同浓度松香酸（20，30，40，50，60，100 μg·mL^-1）为阳性剂的试验系统进行10次试验，应用流式细胞仪技术在人源性淋巴瘤细胞系U937探究最适阳性浓度，并对糖精、对苯二胺、肉桂醇等8个已知其致敏性的化合物进行验证。结果 40，50 μg·mL^-1松香酸能在较小的细胞毒性条件下，表现出稳定的阳性反应，并能对8个已知其致敏性的化合物能进行正确分类。结论 以松香酸为阳性剂建立U-SENS™试验系统可行，可选用终浓度40 μg·mL^-1或50 μg·mL^-1作为阳性对照浓度，确保该试验系统能对受试物的皮肤致敏性进行准确分类。

Abstract:

OBJECTIVE To establish the U-SENS^™ test method which is in line with the actual situation of China, and verify the reliability of the new test method. METHODS Refer to U-SENS™ test method in OECD guideline 442E Appendix II, a test system with different concentrations of abietic acid(20, 30, 40, 50, 60, 100 μg·mL^-1) as the positive agent was established for 10 tests according to the QC template, the optimum positive concentration of human lymphoma cell line U937 was explored by flow cytometry. And eight compounds with known sensitization, such as saccharin, p-phenylenediamine and cinnamyl alcohol were verified. RESULTS The 40, 50 μg·mL^-1 abietic acid could show stable positive reaction under the condition with low cytotoxicity, and could correctly classify 8 compounds with known sensitization. CONCLUSION U-SENS^™ test system is established with abietic acid as positive agent, and 40 or 50 μg·mL^-1 can be chosen as the positive control concentration to ensure that the test system can accurately classify the skin sensitization of the test object.