[Study on feasibility of HaCaT epidermal model as an alternative to skin irritation in vitro].

To construct the epidermal model with HaCaT cells and evaluate the feasibility of this model as an in vitro skin irritation test tool.The HaCaT model was reconstructed with HaCaT cells by adoption gas-liquid surface culture technique, and the EpiKutis ® model was reconstructed with human epidermal keratinocytes by the same techinique as control. Morphology changes of HaCaT and EpiKutis ® models were observed by HE staining. Barrier function assessment was performed with penetration test. According to Organization for Economic Cooperation and Development (OECD) Test Guideline 439, the surface of the HaCaT and the EpiKutis ® models were treated with 20 chemicals for 30 minutes, incubated for 42 hours, and determined tissue viability by MTT assay, to evaluate the irritation of chemicals. Then the results were compared with the irritation of chemicals with the United Nations Globally Harmonized System of Classification and Labelling of Chemicals (UN GHS) system and validated reference method (VRM) for the classification of chemical, and evaluated the feasibility of this model as an in vitro skin irritation test tool.The results of HE staining showed that there was no complete stratified structure in the HaCaT model. The results of barrier function showed that the ET 50 was 0.99 hours. The results of skin irritation of chemicals showed that the sensitivity was 100% and 100%, the specificity was 50% and 70%, and the accuracy rate was 75% and 85% for HaCaT model and EpiKutis ® model respectively.The epidermal model of HaCaT cells does not possess the complete epidermal physiological structure, the barrier function as ET 50 of the HaCaT model is lower than EpiKutis ® model, the chemicals in vitro skin irritation test results do not meet the OECD criteria for the determination of stimulants, so the HaCaT model is not suitable as a replacement tool in vitro to determine the chemicals skin irritation.采用人永生化上皮角质形成细胞系 HaCaT 构建表皮模型，探讨其作为皮肤刺激性替代实验的可行性。.取 HaCaT 细胞采用气-液面培养的方式构建表皮模型，同法以采用人原代上皮角质形成细胞构建的表皮模型 EpiKutis ®作为对照。HE 染色观察两种模型的组织形态；采用快速渗透实验方法检测两种模型屏障功能（ET 50 值）；取 20 种化学标准品分别于两种模型表面受试 30 min，孵育 42 h 后 MTT 法测定组织相对活力百分比，判断化学品刺激性，并与联合国-化学品分类及标记全球协调制度（UN GHS）体内测试结果分类和经济合作与发展组织（OECD）TG439（2013版） 中采用的体外替代方法（validated reference method，VRM）体外测试结果分类进行比较。.HE 染色示，构建的 HaCaT 模型无完整复层化结构；屏障功能检测结果示，ET 50 值为 0.99 h；通过 20 种化学品检测，HaCaT 模型的灵敏度为 100%，特异性为 50%，准确率为 75%；EpiKutis ®模型灵敏度为 100%，特异性为 70%，准确率为 85%。.HaCaT 细胞构建的表皮模型不具备完整的表皮皮肤生理结构特征，屏障功能较低，化学品体外皮肤刺激性测试结果不能完全满足 OECD 关于刺激物的正确判定标准，不适宜作为化学品皮肤刺激性体外替代检测的工具。.