三维细胞支架的制备及在卷烟烟气细胞毒性评价中的应用  

Preparation of three-dimensional cell scaffolds and their application in cytotoxicity evaluation of cigarette smoke

作　　者：李茹洋 尚平平[1] 王宜鹏[1] 贾云祯[1] 孙培建[1] 孙学辉[1] 聂聪[1] 杨松[1] LI Ruyang;SHANG Pingping;WANG Yipeng;JIA Yunzhen;SUN Peijian;SUN Xuehui;NIE Cong;YANG Song(Zhengzhou Tobacco Research Institute of CNTC, Zhengzhou 450001, China)

机构地区：[1]中国烟草总公司郑州烟草研究院

出　　处：《烟草科技》2018年第5期39-45,共7页Tobacco Science & Technology

基　　金：国家自然科学基金项目"高内相乳液法制备聚甲基丙烯酸缩水甘油酯互通多孔材料及其应用研究"(51003122);中国烟草总公司郑州烟草研究院院长科技发展基金项目"一种三维细胞支架的构建及在烟气体外毒理学评价中的应用"(332016CA0210)

摘　　要：为开发用于卷烟烟气体外毒理学评价的三维细胞支架,采用高内相乳液法制备材料,以其为载体进行人肺腺癌细胞A549的三维培养,评价3R4F和CM8两种参比卷烟烟气总粒相物(TPM)的细胞毒性,并与二维平板培养细胞的实验结果进行了对比。结果表明:(1)A549细胞在三维细胞支架上可以吸附、增殖并融合形成细胞层,且在亲水改性支架上增殖更快;(2)培养21d后,三维细胞支架上A549细胞的吸光度(OD值)为3.601,高于二维平板上细胞长满(培养7 d)时的OD值(3.055);(3)3R4F和CM8卷烟烟气TPM浓度与二维平板和三维细胞支架培养的A549细胞存活率均具有良好的剂量-反应关系,且在高TPM浓度时,三维细胞体系下的细胞存活率高于二维细胞体系。该研究为卷烟烟气体外毒理学评价提供了一种新的三维细胞培养方法。In order to develop three-dimensional cell scaffolds for in vitro toxicological evaluation of cigarette smoke, a high internal phase emulsion method was applied to prepare the scaffolding material. Human lung adenocarcinoma cells（A549 cells） were cultured on the scaffolds and then used for the cytotoxicity evaluation of total particulate matter（TPM） in mainstream smoke from two different cigarettes. The evaluation results were compared with those of two-dimensional cell culture systems. The results showed that：1） A549 cells could proliferate and form a cell layer on three-dimensional cell scaffolds. The cells proliferated faster on the scaffolds modified by acrylic acid（Aa）.2） After culturing for 21 days, the optical density（OD） value of A549 cells cultured on the modified three-dimensional cell scaffolds was 3.601, which was higher than that（3.055） of the cells cultured on a two-dimensional plate for 7 days.3） The TPM in smoke of 3R4F and CM8 cigarettes had a good dose-effect relationship with the viability of A549 cells cultured on two-dimensional plate or threedimensional cell scaffolds. At high TPM concentrations, the cell viability was higher on the three-dimensional cell scaffolds than on the two-dimensional plate. This study provides a new three-dimensional cell culture method for in vitro toxicology evaluation of cigarette smoke.

关 键 词：三维细胞支架 二维平板细胞 高内相乳液 人肺腺癌细胞 卷烟烟气 细胞毒性评价 

分 类 号：TS416[农业科学—烟草工业]