兔角膜上皮细胞短时暴露试验的影响因素  

Influencing Factors of Short-term Exposure Test for Rabbit Corneal Epithelial Cells

作　　者：石莹[1] 宋瑞霞[1] 阮鸿洁[1] 张宏伟[1] SHI Ying;SONG Ruixia;RUAN Hongjie;ZHANG Hongwei

机构地区：[1]中国疾病预防控制中心环境与健康相关产品安全所

出　　处：《环境卫生学杂志》2019年第5期489-493,共5页JOURNAL OF ENVIRONMENTAL HYGIENE

摘　　要：目的观察兔角膜上皮细胞短时暴露试验(short-term exposure test,STE test)的影响因素。方法兔角膜上皮细胞分别按3×10^3和6×10^3细胞/孔的初始密度接种,苯扎氯胺等12种化学物质分别以5%和0.05%剂量对细胞染毒5 min后,采用MTT法和CCK-8法显色,计算细胞存活率,采用传统法和STE积分法对上述化学物质的眼刺激性进行评价。结果以3×10^3细胞/孔和6×10^3细胞/孔两种密度接种的细胞,染毒后细胞存活率结果相关性显著。MTT和CCK-8两种显色法测定的细胞存活率结果相关性显著。传统法和STE积分法两种判定标准对重度眼刺激性物质和轻微眼刺激性物质分级判断结果一致。结论细胞初始接种密度、不同显色方法和结果判定标准是兔角膜上皮细胞短时暴露试验结果的影响因素,可采用3×10^3细胞/孔或6×10^3细胞/孔作为试验初始接种密度,采用CCK-8法测定细胞活性,STE积分法判定受试物对眼的刺激强度。Objectives To observe the influencing factors of short-term exposure test(STE test)in rabbit corneal epithelial cells.Methods Statens Seruminstitut Rabbit Cornea cells were seeded at the initial density of 3×10^3 and 6×10^3 cells/well respectively.12 chemicals,including benzalkonium chloride,were exposed to cultured cells at the doses of 5%and 0.05%respectively for 5 minutes,and cell viability were detected by MTT assay and cck-8 assay,The eye irritation of the above chemicals was evaluated by the traditional method and the STE integration method.Results Seeded at 3×10^3 cells/well and 6×10^3 cells/well,the cell viability result were correlated well after exposure.The cell viability result measured by MTT assay and CCK-8 assay were significantly correlated.The traditional method and STE integration method were consistented with the result of grading judgments for severely irritating substances and mild irritants.Conclusions The initial cell inoculation density,different color method and result criteria were the influencing factors of short-term exposure test of rabbit corneal epithelial cells.The density of 3×10^3 or 6×10^3 cells/well,the CCK-8 assay for cell viability determination,and the STE integration method for the eye irritation evaluation could be optional for the experiment.

关 键 词：眼刺激性 兔角膜上皮细胞 显色法 

分 类 号：R114[医药卫生—卫生毒理学]