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日用化学工业 ›› 2018, Vol. 48 ›› Issue (5): 288-292.doi: 10.13218/j.cnki.csdc.2018.05.009

• 开发与应用 • 上一篇    下一篇

基于皮肤模型与THP-1细胞共培养建立皮肤致敏体外检测方法的初步研究

陈健1,2, 柯逸晖1,2, 谈伟君1   

  1. 1.广东药科大学,广东 广州 510623;
    2.广州市华代生物科技有限公司,广东 广州 510623
  • 收稿日期:2017-12-29 发布日期:2019-03-18
  • 通讯作者: 谈伟君(1963-),男,教授,E-mail:weijunt0726@163.com。
  • 作者简介:陈 健(1990-),男,硕士研究生,E-mail:chenjian1238@126.com。
  • 基金资助:
    广东省科技计划项目(2015A030402005)

A preliminary study on in vitro skin sensitization test based on skin model and THP-1 cell co-culture

CHAN Jian1,2, KE Yi-hui1,2, TAN Wei-jun1   

  1. 1.Guangdong Pharmaceutical University,Guangzhou,Guangdong 510623,China;
    2.Guangzhou Chn-Alt Biotech.Co.,Ltd.,Guangzhou,Guangdong 510623,China
  • Received:2017-12-29 Published:2019-03-18

摘要: 建立了皮肤模型与THP-1细胞共培养方法,对石化产品进行了致敏性评价。不同浓度测试物与表皮模型作用15 min,去除受试物后孵育6 h,使用MTT法检测皮肤模型组织活率,利用流式细胞法检测THP-1细胞表面标志物CD86、CD54表达变化情况以及细胞活率。对上述参数进行统计学分析及是否存在剂量效应关系,判定测试物是否具有潜在致敏特性。对SLS、肉桂醛、柠檬醛的测试表明,该方法可以正确区分刺激物和致敏物;对4种未知润滑油添加剂的预测结果表明,HVI60、Cuvan826、ATM PAN为非致敏物,Irgamet30为致敏物,高浓度时可导致表皮组织和THP-1细胞活性降低及表面标志物CD86、CD54的特异性升高。

关键词: 皮肤致敏, 皮肤模型, THP-1细胞, 共培养

Abstract: An in vitro skin sensitization test based on skin model and THP-1 cell co-culture was established,and used to evaluate the sensitizing potency of petrochemicals.Skin model was treated with different concentrations of samples for 15 min,then removing the samples and was incubated for another 6 h.Skin tissue activity was measured by MTT assay,while cell viability and cell surface markers (CD54,CD86) expression levels of THP-1 cells were detected by flow cytometry.The skin sensitization potential of test substances was determined by statistically analyzing the above parameters and reaching a dose-response relationship.The results of SLS,cinnamaldehyde and citral show that this co-culture method can correctly distinguish irritants and sensitizers.The results of 4 unknown lubricant additives show that HVI60,Cuvan826 and ATM PAN are predicted to be non-sensitizers,while Irgamet30 is predicted to be a sensitizer,which at higher concentration can decrease the skin tissue activity and THP-1 cell viability,and specifically increase the expression of surface markers.

Key words: skin sensitization, skin model, THP-1 cells, co-culture

中图分类号: 

  • R99