用于肿瘤细胞三维培养的微流控芯片构建及培养条件研究

摘要

目的：设计制作用于肿瘤细胞三维培养的微流控芯片,以海藻酸钙凝胶为细胞支架,基于芯片研究适合的凝胶形成、溶解条件,利用该条件长期培养SMMC-7721细胞并检测其活力。方法利用CDR软件、软光刻技术、模塑法、等离子键合法等设计并制作出微流控芯片并验证其适用性；在芯片中三维培养肝肿瘤细胞SMMC-7721,观察细胞形态,以IPP软件辅助计算细胞在72h的存活率。结果设计制作的微流控芯片适合于肿瘤细胞三维培养；芯片中培养的肝癌细胞72 h内生长状态良好,存活率达(96．1±4．5)%,细胞堆积排列紧密,出现肿瘤细胞聚集体( tumor cell spheroid, TCS)。结论该文设计构建用于肿瘤细胞三维培养的微流控芯片,并用于肝癌细胞SMMC-7721培养,细胞生长状态良好,且细胞生长与聚集状态表现出一定特殊性。利用该芯片进行肿瘤细胞三维培养,可以为肿瘤细胞生长状态研究及抗肿瘤药物筛选提供更有利的方式。%Aims To design and fabricate the 3 D cell cultural microfluidic chip for tumor cell culturing, with which to research the compatible conditions for gelatin forming and dissolving with calcium alginate as the scaffolds. To culture SMMC-7721 cells in the chip and to detect the surviving rate. Methods The microfluid-ic chip was fabricated with the software Corel Draw, the technology of soft lithography, molding, and plas-ma bonding. The applicability was tested and cells were cultured on it, on which the cell status was ob-served, their surviving rate was calculated with the help of the software IPP. Results The chip we fabri-cate was calculated is suitable for cell 3D culturing, the tumor cells showed a favorable proliferation ability in 72 h on chip, the surviving rate was ( 96. 1 ± 4. 5)%. The cells were solid and TCS appeared. Con-clusion The microfluidic chip manufactured appeared for tumor cell 3D culture, is suitable for the growth of SMMC-7721 and the cells are indubitable. They show some different status in proliferative and agminated compared with traditional 2D cell culture. With the chip and the condition found, there will be a better way to study the characteristics of tumor cells and is beneficial to the screen of anti-tumor drugs.