Hoh A, Maier K, Dreher R M
Battelle Europe, Department of Molecular and Cellular Biology, Frankfurt, Federal Republic of Germany.
Mol Toxicol. 1987;1(4):537-46.
Human keratinocytes are the most appropriate target cells for evaluating mechanisms of skin cytotoxicity and pharmacology of chemical agents. After having formed a confluent stratified epithelium with proliferating basal cells and differentiated cell layers, human keratinocytes were harvested after enzymatic detachment as stable, three-dimensional cell aggregates or used as adherent multilayers in microtiter plates to study the local cytotoxicity of different toxic compounds. The advantage of this test is that it uses the adequate target cells and that it evaluates both the ability of the test chemical to penetrate several cellular layers as well as the ability to interfere with cellular function. The end points are cell viability and cell metabolism, which are determined by neutral red uptake and MTT reduction, respectively. For the 13 chemicals evaluated in this study we found good correlation (r = .819) between the potency rankings of keratinocyte NR 50 values and in vivo irritancy data. There was also good agreement (r = .945) between ranking of these chemicals according to midpoint toxicity of both the 3T3 assay and the keratinocyte assay. This test system might be at the present stage a supplementation of the current test battery, which shall replace in vivo irritation tests like the Draize test.
人类角质形成细胞是评估皮肤细胞毒性机制和化学物质药理学的最合适靶细胞。在形成具有增殖性基底细胞和分化细胞层的汇合分层上皮后，通过酶解分离收获人类角质形成细胞，作为稳定的三维细胞聚集体，或用作微孔板中的贴壁多层细胞，以研究不同有毒化合物的局部细胞毒性。该试验的优点在于它使用了合适的靶细胞，并且它既评估了受试化学物质穿透多个细胞层的能力，也评估了其干扰细胞功能的能力。终点指标是细胞活力和细胞代谢，分别通过中性红摄取和MTT还原测定。对于本研究中评估的13种化学物质，我们发现角质形成细胞NR 50值的效力排名与体内刺激性数据之间具有良好的相关性（r = 0.819）。根据3T3试验和角质形成细胞试验的中点毒性对这些化学物质进行的排名之间也具有良好的一致性（r = 0.945）。该试验系统在现阶段可能是对当前试验组合的一种补充，有望取代像Draize试验这样的体内刺激性试验。
