肺腺癌细胞株体外三维培养模型的建立及其药物敏感性的观察

Construction of three dimensional in vitro culture model of pulmonary adenocarcinoma and detection of its drug resistance

目的 探讨肿瘤细胞三维培养的理想方法 ,为模拟体内实体瘤细胞间相互作用提供良好模型。方法 在抑制细胞贴壁、间断振荡条件下 ,培养无载体三维细胞模型 ;利用微载体CutiSpher进行多细胞球 (multicellularspheroids ,MCS)培养 ;MTT法检测单层细胞 (monolayercells ,MC)药物敏感性 ,细胞计数法检测MCS药物敏感性 ;利用透射电镜比较MCS与MC超微结构的差异。结果 无载体三维细胞培养在培养 4d后即可形成由多个细胞组成的细胞团。MCS在培养 0 5h后即可见细胞贴附于载体 ;培养 5d后 ,形成有多层细胞结构的细胞球。与MC相比 ,MCS药物敏感性显著下降 ;细胞器及微绒毛丰富 ;细胞间粘附广泛而紧密。结论 MCS细胞生长活性好 ,能较好地模拟体内实体瘤的细胞间相互作用 ,特别是细胞间粘附 ;

Objective To explore an optimal model of three dimensional in vitro cell culture for simulating solid tumors in vivo . Methods The model of three dimensional cell culture was constructed under the conditions of inhibiting the cell wall attachment and stirring the medium. Multicellular spheroids (MCS) were cultured using microcarriers (CutiSpher). Drug sensitivity of monolayer cells (MC) and MCS was tested by MTT staining and cytometry, respectively. Ultrastructures of the MC and MCS were observed by transmission electron microscopy. Results Cells in three dimensional cell culture model without microcarriers were compacted into mass at 4 d while cells in MCS were found to attach to the microcarriers at 0.5 h. MCS had more than two layers of cells growing within it at 5 d. Compared with MC, MCS was more resistant to the anticancer drug, and had more plenty of organell and microvilli with more extensive and compact cell adhesion. Conclusion MCS has strong developmental properties and can simulate the cell cell interactions in vivo , especially cell adhesion, which may contribute to the drug resistance of MCS.