体外琼脂糖三维培养肺腺癌细胞株A549的实验研究  

Investigation on Establishment of Three-dimensional Cell Culture Model of A549 with Agarose in Vitro

作　　者：章洪华 吴智春[2] 欧阳兵 季旭明 ZHANG Honghua;WU Zhichun;OUYANG Bing;JI Xuming(Shandong University of Traditional Chinese Medicine, Jinan 250355, Shandong, China;Jining Medical College, Jining 272067, Shandong, China;Shandong Food and Drug Administration, Jinan 250011, Shandong, China;Shandong Classic formula engineering demonstration center, Jinan 250355, Shandong, China)

机构地区：[1]山东中医药大学,山东济南250355 [2]济宁医学院,山东济宁272067 [3]山东食品药品监督管理局,山东济南250011 [4]山东省经方工程示范中心,山东济南250355

出　　处：《中华中医药学刊》2018年第4期827-829,I0005,共4页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81573871,81072832,81273634);山东省重点产业关键技术创新项目(2016CYJS08A01-3,2016CYJS08A01-4);山东省重点研发计划项目(2016ZDJS07A12)

摘　　要：目的：体外研究琼脂糖用于肺腺癌细胞株A549三维培养,并观察细胞在三维环境中的行为改变。方法：用琼脂糖作为三维培养基质,观察细胞在三维环境中的生长现象;黏附实验用以检测细胞对胞外基质的黏附率;MTT比色法检测细胞对DDP的药物敏感性和细胞增殖抑制率;流式细胞术检测细胞自然凋亡情况。同时比较二维及三维培养细胞的差异。结果：在琼脂糖中,A549细胞生长呈现球形的团块状生长,细胞团块大小不一,生长状态良好。随着细胞的增长,3D培养的A549细胞对DDP的抗药性增加,药敏性降低;二维和三维培养下的A549细胞黏附率存在显著差异;3D培养下的细胞自然凋亡明显减少。结论：A549细胞在琼脂糖三维培养中,其细胞行为与二维培养时发生显著改变,二者具有显著性的差异。因此,琼脂糖可用于构建A549三维培养模型,此模型可为进一步研究提供研究基础。Objective： We aims to establish the three-dimensional culture model of lung cancer cell line A549 with agarose and study the cellular behaviors of these cells. Methods： We established three-dimensional culture model of A549 cell line with agarose. The morphology,distribution and viability of A549 cells in three-dimensional culture were identified by microscopy. MTT assay was used to assess cisplatin cytotoxicity to the cell colonies. The adhesion abilities of cells of three-dimensional culture and two dimensional culture were measured by their adhesion percentage to ECM proteins. Flow cytometry was applied to study the influence of A549 cells＇ apoptosis. Results： Agarose formed block mass and could be used as culture matrix to mimic the cell growth conditions in vivo and the three-dimensional culture for cells in vitro. In this study,cells grown in Agarose formed multicellular colonies and good viability. The cell resistance to cisplatin increased when cells formed multicelluar colonies following the culture time and reduced the sensibility of cells to DDP. The adhesion assay indicated that there were significant differences of cells in three-dimensional culture to ECM proteins compared to two-dimensional culture. Flow cytometry showed us the A549 cells＇ apoptosis decline in three-dimensional cell culture. Conclusion： Our present study showed that three-dimensional culture of A549 cells in agarose matrix influenced the cellular behaviors,including the cell growth,morphology,drug resistance alteration and the adhesion ability,which were significantly different from cells in two-dimensional culture. Three-dimensional culture constructed using agarose can provide a significant research basis for further study in the future.

关 键 词：琼脂糖 三维培养 肺腺癌细胞 

分 类 号：R285.5[医药卫生—中药学]