Table 1.
The advantages and disadvantages of methods used for migration/invasion study
| Method | Advantages | Disadvantages | ||
|---|---|---|---|---|
| 2D simple methods (migration) | Lack of tumor microenvironment | Time-lapse microscopy | Very simple method | Only for individual cells; accuracy depends on software analysis |
| Wound healing assay (scratch assay) | Easy, quick, cheap, repeatable | Cell stress and retraction of the edges, impact of proliferation, lower reproducibility | ||
| Barrier assay | Easy, quick, cheap, repeatable; decreased cell stress and retraction; higher reproducibility | Impact of proliferation | ||
| Scatter test | Morphological changes and movement in response to HGF | Now not widely used | ||
| 2D chemotactic method (migration) | Dunn chamber | Directed chemotactic cellular movement | Movement of individual cells is assessed | |
| Videomicroscopy of cells | Live monitoring of cell movement; assessing of various movement parameters | Instrumentation is needed: camera connected to microscope and incubation chamber | ||
| Chemoattractive beads | Movement of cell sets; comparison of different cell population; ex vivo explant analysis | Now not widely used | ||
| 2D/3D Boyden chamber (migration/invasion) | Boyden chamber | Cellular phenotype closer to in vivo conditions; active movement of cells through membrane | Process of cell movement is not visualized | |
| Transwell | Homogenous multiwell platform: different conditions, various cell types or assay settings may be included in one plate; commercially available; high sensitivity to chemoattractant | Limited time of performance | ||
| xCELLigence | On-line monitoring of cells (also including information about proliferation and adhesion); multiwell setting | Special instrument and plates needed | ||
| in vitro 3D methods | 3D microscopy | Various microscope types according to type of experiment; live imaging—movement of cells in space and time; monitoring of subcellular structures as well as whole organ/organism | Expensive and highly sophisticated instruments; special cultivation plastics; expertized staff | |
| Microfluidic devices | Movement of cells in the fluid flow mimics body fluidics; customized precisely defined platforms; combination of various environment parameters (cellular, molecular, chemical, biophysical) in one assay; suitable for 3D cell structures | Special instrumentation controlling microflow miniaturized platforms—worse handling; expertized staff | ||
| 3D cultures | Cellular phenotype closer to in vivo conditions various type of 3D cell structures; co-cultivation of various cell types (crosstalk); commercial multiwell platform; human tumor cultivation to improve personalized medicine (patient´s consent); state-of-the-art techniques comparable to in vivo mice studies | Long-term manipulation; special cultivation media and plastics; 3D fluorescence microscopy is necessary | ||
| in vivo methods | In vivo mice studies | Crosstalk of cancer cell and organism including immune cell in some cases; large offer of various mice strains for various purposes; transplant models of human tumors; therapy testing | Long-term duration of experiments; time-consuming, expensive; special laboratories needed; results not always transferable to human; ethical controversies | |