Matrigel胶的肺腺癌细胞株A549体外三维培养的实验研究

Three dimensional culture model of lung cancer cell line A549 and the study of their cellular behaviors

目的研究肺腺癌细胞株A549三维体系培养及其细胞行为的改变。方法对三维培养基质Matrigel水凝胶进行材料表征,研究其纳米纤维网络的形成。观察细胞在三维环境中的克隆形成,形貌及存活力,并检测了细胞形成克隆后的药敏性,同时比较二维及三维培养的细胞对胞外基质蛋白粘附率的差异。结果Matrigel胶形成了类似于体内胞外基质的纳米纤维网络结构。在Matrigel胶中,A549细胞生长呈现出球形的多细胞克隆,生长状态均较好,与材料具有较好的生物相容性。随着克隆的增长,细胞对外界刺激的抵抗性及抗药性增加,药敏性降低。二维和三维培养下的A549细胞对不同的胞外基质蛋白的粘附率存在显著差异。结论细胞在Matrigel胶三维培养中,细胞的生长,形貌,存活力,药敏性及粘附性等细胞行为与二维培养的细胞具有显著性的差异,可为抗药性研究及细胞信号通路的研究等提供研究基础。

Objective Our aims to establish the three dimensional culture model of lung cancer cell line A549 and to study the cellular behaviors of these cells. Methods Transmission Electron Microscopy results demonstrated the nanofiber networks formed by the Matrigel hydrogel. The morphology, distribution and viability of A549 cells in three dimensional culture were identified by fluorescence microscopy. MTT assay was used to assess 5-Fluorouracil cytotoxicity to the cell colonies. The adhesion ability of cells of three dimensional culture and twodimensional culture was measured by their adhesion percentage to ECM proteins. Results Matrigel formed nanofiber networks 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 Matrigel formed multieellular colonies and good viability and bioeompatibility with the matrix. The cell resistance to 5 Fluorouracil increased when cells formed multicelluar colonies following the culture time and reduced the sensibility of cells to the drug. The adhesion assay indicated that there were significant differences of cells in three dimensional culture to ECM proteins compared to two dimensional culture. Conclusion Our present study showed that three dimensional culture of A549 cells in matrigel matrix influenced the cellular behaviors including the cell growth, morphology, viability, drug resistance alteration and the adhesion ability which were significantly different from cells in two dimensional culture. Three dimensional culture can provide significant research basis for the study of drug resistance and the cell signaling pathways in the future.