丝裂霉素C作为琼脂扩散法细胞毒性试验阳性对照材料的探讨

Study on the Applicability of Mytomycin C as a Positive Control Material for Agar Diffusion Cytotoxicity Test

目的为细胞毒性评价方法——琼脂扩散法探寻具有适度细胞毒性、挥发性小且易于获取的阳性对照材料。方法按照ISO10993.5-2009与YY/T 0127.9-2001规定的琼脂扩散法,使用L-929细胞接种于培养皿内,培养至近汇合单层细胞,倒入琼脂并用中性红溶液染色,丝裂霉素C作为试验材料进行试验,样品浓度分别为2 mg/m L、1 mg/m L与0.5 mg/m L。结果 2 mg/m L丝裂霉素C组毒性分级为3级;1 mg/m L丝裂霉素C组毒性分级为3级;0.5 mg/m L丝裂霉素C组毒性分级为2级。结论 2 mg/m L浓度的丝裂霉素C可作为琼脂扩散法评价细胞毒性时的阳性对照材料。

Objective To find a non-volatile positive material control which showed moderately or severely cytotoxic and could be obtained easily. Methods The in vitro cytotoxicity study was conducted according to the agar diffusion method in ISO 10993.5-2009 and YY/T 0127.9-2001. Each sample was placed in contacted with the surface of an agar layer overlaying a monolayer of L-929 Mouse Fibroblast cells which were stained with neutral red vital stain at the bottom of the plate. As the sample, Mytomycin C was diluted to 2 mg/mL, 1 mg/m L and 0.5 mg/mL. Results In the group of 2 mg/m L Mytomycin C, it was observed that the cytotoxicity grade was 3. In the group of 1 mg/m L Mytomycin C, it was observed that the cytotoxicity grade was 3. In the group of 0.5 mg/mL Mytomycin C, it was observed that the cytotoxicity grade was 2. Conclusion 2 mg/mL Mytomycin C could be used as a positive material control of agar diffusion.