Validation Study of the In Vitro Skin Irritation Test with the LabCyte EPI-MODEL24
Validation Study of the In Vitro Skin Irritation Test with the LabCyte EPI-MODEL24
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DOI：
10.1177/026119291204000108
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发表时间：
2012-03-01
影响因子：
2.7
通讯作者：
Omori, Takashi
中科院分区：
文献类型：
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作者：
Kojima, Hajime;Ando, Yoko;Omori, Takashi
使用日本组织工程有限公司（Gamagori，Japan）开发的重建人表皮（RhE）LabCyte EPI-MODEL 24进行体外皮肤刺激试验的验证研究。本研究中遵循的方案是EpiSkin方案（LabCyte测定）的优化版本。根据评估化学品皮肤刺激潜力的联合国全球协调制度（UN GHS）分类，由日本动物实验替代品协会（JSAAE）皮肤刺激试验验证研究管理团队（VMT）的至少6个实验室对12种刺激物和13种非刺激物进行了验证。这25种化学品被列入欧洲替代方法验证中心（ECVAM）的性能标准。将重建的组织暴露于化学品15分钟，并在新鲜培养基中孵育42小时。随后，测量条件培养基中存在的白细胞介素-1 α（IL-1 α）水平，并使用MU测定法评估组织活力。采用LabCyte EPI-MODEL 24（LabCyte MU检测试剂盒）获得的MIT检测试剂盒结果证明，实验室内和实验室间重现性较高，并且作为独立检测试剂盒用于区分皮肤刺激物和非刺激物的准确度较高。此外，LabCyte试验中的IL-1 α释放测量对于该模型在皮肤刺激潜力化学品分类中的成功显然是不必要的。
A validation study on an in vitro skin irritation assay was performed with the reconstructed human epidermis (RhE) LabCyte EPI-MODEL24, developed by Japan Tissue Engineering Co. Ltd (Gamagori, Japan). The protocol that was followed in the current study was an optimised version of the EpiSkin protocol (LabCyte assay). According to the United Nations Globally Harmonised System (UN GHS) of classification for assessing the skin irritation potential of a chemical, 12 irritants and 13 non-irritants were validated by a minimum of six laboratories from the Japanese Society for Alternatives to Animal Experiments (JSAAE) skin irritation assay validation study management team (VMT). The 25 chemicals were listed in the European Centre for the Validation of Alternative Methods (ECVAM) performance standards. The reconstructed tissues were exposed to the chemicals for 15 minutes and incubated for 42 hours in fresh culture medium. Subsequently, the level of interleukin-1 alpha (IL-1 alpha) present in the conditioned medium was measured, and tissue viability was assessed by using the MU assay. The results of the MIT assay obtained with the LabCyte EPI-MODEL24 (LabCyte MU assay) demonstrated high within-laboratory and between-laboratory reproducibility, as well as high accuracy for use as a stand-alone assay to distinguish skin irritants from non-irritants. In addition, the IL-1 alpha release measurements in the LabCyte assay were clearly unnecessary for the success of this model in the classification of chemicals for skin irritation potential.