The primary study on the application of gelatum invasive model as an invasiveness assay for glioma cells in vitro

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. DOI：10.3969/j.issn.1672-6731.2010.05.00