Abstract: Objective To investigate the feasibility of applying gelatum invasive model as an invasiveness assay for human glioma cells in vitro, observe the differences of invasiveness between adherently cultured differentiated human glioma cells and human glioma stem cells (GSCs), and validate it as a novel method for in vitro glioma invasiveness research. Methods The spheres [(10-15) × 10 3 cells/ sample] formed by hanging drops from primary cultured adherent glioma cells and glioma stem cells spheres were planted respectively into gelatum invasive model and cultured in the corresponding medium with matrix metalloproteinases (MMPs) inhibitor GM6001 at different concentrations (0, 25, 50, 75, 100 μmol/L) in 37 ℃, 5% CO2. The distance of cells migration in the 2 groups was recorded by microscope (× 240) and measured through Photoshop software, and then analysed by SPSS 16.0. Results The initial adherent cells spheres showed spherical and cells grew well in gelatum invasive model, and invaded in the gelatum with time. The inhibitory effects of GM6001 on invasiveness of the 2 groups were distinctive and showed a dose-dependent manner compared with the controls. GM6001 at 75 μ mol/L had the greatest inhibitory effect on human glioma differentiated cells (P = 0.000, for all) after 4 d culture compared with the controls, whereas for human glioma stem cells spheres it was at 25 μmol/L (P = 0.002, 0.012, 0.000). Conclusion The gelatum invasive model represents a useful and valid invasiveness assay which is applicable to both adherently cultured glioma cells and glioma stem cells spheres.

Key words: Glioma, Neoplasm invasiveness, Collagen, Matrix metalloproteinases, Cells, cultured

摘要： 目的   初步探讨凝胶侵袭模型用于人胶质瘤细胞体外侵袭力研究的可行性，观察普通培养的胶质瘤细胞与胶质瘤干细胞在凝胶侵袭模型中侵袭力的差异，为胶质瘤的体外研究寻求新的实验手段。方法   经纯化的Ⅰ型和Ⅲ型胶原与MEM 培养基混合制备凝胶液，将原代培养的胶质瘤细胞经悬滴法制成的细胞球和胶质瘤干细胞所形成的细胞球种植于其中，每个细胞球含（10 ~ 15）× 103 个细胞，在添加含有不同药物浓度基质金属蛋白酶抑制剂GM6001（0、25、50、75、100 μmol/L）的相应培养基中培养4 d，测量不同药物浓度组肿瘤细胞侵袭距离及其差异。结果   不同处理组胶质瘤细胞在凝胶侵袭模型中生长良好，最初的细胞球形态呈规则圆形，随着时间的推移肿瘤细胞侵袭距离逐渐增加；且对GM6001 的抑制作用表现出不同程度的剂量依赖性，以75 μmol/L GM6001 对普通培养的胶质瘤贴壁细胞的侵袭抑制作用最为明显（均P = 0.000）；而25 μmol/L GM6001 对胶质瘤干细胞球的侵袭抑制作用最显著（P = 0.002，0.012，0.000）。结论   凝胶侵袭模型适用于普通贴壁培养的胶质瘤细胞和胶质瘤干细胞球。

关键词: 神经胶质瘤, 肿瘤侵润, 胶原, 基质金属蛋白酶类, 细胞, 培养的